EPA 910/9-91-042
Bioaccumulation of Contaminants in Crabs and Clams
in Bellingham Bay
Prepared for
Puget Sound Estuary Program
US Environmental Protection Agency
Region 10, Seattle WA
and
Fran Solomon and Lucy Pebles
Northwest Regional Office
Washington State Department of Ecology
by
James Cubbage
Washington State Department of Ecology
Environmental Investigations and Laboratory Services
Toxics, Compliance, and Ground Water Investigations Section
Olympia, WA 98504-6814
Waterbody No. WA-01-0040
(Segment No. 01-01-02)
September 1991
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ABSTRACT
Bellingham Bay supports commercial and recreational shellfish harvest. Industrial and municipal
discharges into the bay have contributed to high sediment concentrations of mercury and PCBs.
To investigate potential bioaccumulation of contaminants in shellfish, crab (Cancer magister)
muscle was collected from eight sites and littleneck clams (Tapes japonica, Protothaca stamnea)
from four sites and tested for concentrations of PCBs, other organochlorine compounds including
chlorpyrifos and pentachlorophenol, cadmium, arsenic, lead, and-mercury. Clams were also
tested for polycyclic aromatic hydrocarbons (PAH). In crabs, metals ranged as follows:
(Cd:0.002-0.005; As: 1.9-5.6; Pb:0.05-0.29; Hg:0.06-0.15; in mg/kg wet weight). Metals in
whole clams ranged as follows: (Cd:0.18-0.23; As: 1.1-2.0; Pb:0.02-0.15; Hg:not detected-0.02;
all mg/kg wet weight). Low concentrations of PAH were found in clams (<2-20 ppb). No
pesticides or PCB's were found above detection limit in crabs or clams. A concurrent study
conducted by Department of Natural Resources that examined crabs caught from the center of
the bay found equivalent concentrations of mercury, arsenic and lead, higher concentrations of
cadmium and detections of DDE in 2 of 7 samples (1.5, 0.6 ppb) and chlordane in 1 of 7
samples (3.7 ppb). These metals and PAH concentrations are comparable to concentrations
found in Puget Sound reference areas with presumably low levels of contamination.
Concentrations of mercury in Bellingham Bay crabs have declined over the last 15 years and
reflect the decreased discharge of mercury into the bay.
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ACKNOWLEDGEMENTS
Several people and agencies contributed to this study. This project was conducted cooperatively
with a Department of Natural Resources (DNR) study to determine chemical concentrations in
biota at Puget Sound Dredge Disposal Analysis (PSDDA) sites within Bellingham Bay. This
cooperation saved logistic costs and provided greater spacial coverage. Mike MacKay of the
Lummi Tribe collected crabs with crab pots and assisted in study design. Michael Cochrane of
the Lummi Tribe assisted in collecting crabs. SAIC under contract to DNR provided assistance
in the field and the use of the R. V. Kittiwake. Charlie Eaton provided and piloted the R. V.
Kittiwake. Sample analysis was supervised by Dick Huntamer, Bob Rieck, Stuart Magoon and
Craig Smith of the Manchester Environmental Laboratory. Advice and review of the sampling
plan was provided by Betsy Striplin of DNR, Quality Assurance section of the Environmental
Protection Agency (EPA) and the Department of Ecology, and Dr. Jacques Faigenblum of U.S.
EPA. Project oversight was provided by Dr. Fran Solomon and Lucy Pebles of Department of
Ecology. This report was critically reviewed by Art Johnson, Betsy Striplin, and Dr. Fran
Solomon. Kelly Carruth and Gayla Diamond typeset and proofread this manuscript. Funding
was provided by the U. S. EPA, Puget Sound Estuary Program. Dave Smith administered the
contract. I would like to thank all these people and organizations.
11
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INTRODUCTION
Evidence suggests several kinds of contaminants that bioaccumulate occur or could occur in
Bellingham Bay biota. Mercury and PCBs have been found in Bellingham Bay sediments (PTI,
1989) at concentrations that exceed sediment quality criteria and thus may be harming marine
biota. Pentachlorophenol (PCP) has been found in sediments in Whatcom Creek, a stream that
empties into Bellingham Bay (Kendra, 1987). These chemicals can bioaccumulate. Chlorpyrifos
is a pesticide of concern in Puget Sound due to its patterns of use and persistence through
trophic levels (Tetra Tech, 1988). The Nooksack River, which drains a large agricultural area,
may convey pesticide-laden water or sediments into the bay. Arsenic, cadmium, and lead often
bioaccumulate and are associated with urban areas. Polycyclic aromatic hydrocarbons (PAH)
are potentially harmful compounds often found in relatively high concentrations in urban
sediments. They also bioaccumulate in some species. Bellingham Bay supports commercial and
recreational crab and clam fisheries.
This study, conducted under EPA's Puget Sound Estuary Program, examined edible crab and
clam tissue to determine concentrations of these potential contaminants in the food chain.
Concurrent to this effort was a study conducted by SAIC (1991) for the Washington State
Department of Natural Resources to collect and analyze crab tissues for contaminants. This
work was designed to determine concentrations of contaminants in crabs at the Puget Sound
Dredged Disposal Analysis ( PSDDA) disposal site in the middle of Bellingham Bay as well as
near industrial and rural areas of the bay. The work was also designed to monitor the relative
distribution of crabs within the bay and near the disposal sites.
METHODS
Locations
Figure 1 shows sample collection sites for this study. Table 1 shows the sampling dates, number
of individuals collected, and location of samples. These sites were chosen to both reflect areas
of recreational use and potential areas of contamination. Adult Dungeness crabs (Cancer
magister) were collected at eight sites near the shores of Bellingham Bay. Native littleneck
clams (Protothaca stamined) and Japanese littleneck clams (Tapes japonica) were collected at
four sites.
Collection Methods
In cooperation with the Lummi Tribe, crabs were collected at all sites except Site 4 with
commercial crab pots. These pots were set for 10-18 hours before being hauled and checked.
At Site 4, crabs were captured with a 3 meter beam trawl towed behind the research vessel
Kittiwake at 1.5 kts. Other sites were sampled with the beam trawl, but inadequate numbers of
adult male crabs were recovered. Only crabs legal for commercial and recreational harvest were
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BELLINGHAM BAY
Figure 1. Study area and sampling sites.
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Table 1. Sampling sites for Bellingham Bay bioaccumulation study.
Site
Date
Collected
Dungeness crab
1
2
3
4
5
6
7
8
8/20-21/90
9/05/90
8/20-21/90
8/20/90
9/05/90
8/20-21/90
9/05/90
8/22/90
Samples Recovered(l)
Female
>«.2Sln
Males
Latitude
<6.25ln Collected
Location
Longitude epth
(m)
Sampling site description
Cancer nugister
9
2
1
40
18
7
8
Num
6
6
5
4
3
8
7
5
Av
6 5
3 5
2 5
1 4
2 3
5 5
4 7
5
Size (mm)
Range
48
48
48
48
48
48
48
48
Species
41.3
43.9
44.0
42.5
43.2
43.5
44.1
44.5
122
122
122
122
122
122
122
122
29.4
36.7
30.1
31.2
31.3
30.5,
v
31.0
29.5
33
8
13
20
21
16
15
9
Chuclcanut Bay
East side Lummi Penninsula near Brandt
Off dock in Boulevard Park
Island
Off Post Point marine park (caught with trawls)
Post Point near municipal outfall diffuser
Padden Creek mouth near Fairhaven boat
Buoy at Georgia Pacific diffuser outfall
Mouth of Whatcom Waterway
launch
Littleneck clams
IB
2B
3B
4B
9/05/90
8/20/90
12/14/90
8/20/90
22
30
20
28
51.4
37.1
47.0
45.3
45-58
32-48
40-58
36-54
Ps 48
Tj 48
Ps 48
Tj 48
40.3
44.0
43.7
43.5
122
122
122
122
29.3
36.7
31.0
37.0
Chuclcanut Bay (South side near Yacht Club bay)
East side of Lummi Peninsula
Post Point marine park
Brandt Island
(1) For crabs: number of animals caught in traps. Collected refers to number taken for analysis.
For crabs, only males 76.25 inches were analyzed.
(2) Species: Ps «Prototbtc* sttmiaea
Tj = Tapes japonic*
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sampled (males with carapaces wider than 6.25 inches). For all trapped crabs, sampled animals
were killed with a blow to the ventral surface, wrapped in aluminum foil and frozen up side
down to minimize contamination of muscle by hepato-pancreas fluids. For the trawled crabs,
animals were wrapped alive in foil and frozen upside down.
Clams were collected with shovels and rakes off the beach at low tide. Clams were sampled
from at least two areas at least 20 meters apart. A sample of at least 20 clams was collected,
rinsed with site water, and frozen at the earliest opportunity. The clams were not allowed to
depurate in order to provide a potential worst case exposure to recreational users.
Sample Preparation
All stainless steel sample tools (forceps, scalpel, and knives) and blenders were decontaminated
with the following procedure:
1) Wash in hot water and Alconox detergent;
2) rinse in tap water;
3) rinse in 10% nitric acid;
4) rinse with deionized water;
5) rinse with pesticide analysis grade acetone; and
6) air dried.
Clam sizes and crab carapace widths were measured. Samples from each site were shelled
separately. Muscle from crabs was collected while still partially frozen by breaking off the legs
and claws from the body, cutting the shell with scissors and scooping out the muscle with
stainless steel scalpels. All tissue and fluid from whole shelled clams and fluid were scooped
out with stainless steel spoons. Tissues from 4-7 crabs and 20-30 clams from each site were
homogenized in a decontaminated Waring blender and poured into pollutant-free glass jars with
teflon-lined lids (ICHEM series 300) and frozen. Samples were frozen within 48 hours of
collection and extracted within 35 days of dissection, or, within 60 days of original collection.
Sample Analysis
Samples were analyzed for percent solids, percent lipids, mercury, arsenic, lead, cadmium,
PCBs, chlorinated pesticides and chlorpyrifos. Crab samples were also analyzed for penta-
chlorophenol and its breakdown products. Table 2 presents sample analyses. Clam samples
were also analyzed for PAH. Table 3 reviews the schedule of analyses. Due to field collection
problems, sample 3B was collected three months later than the other samples. It was analyzed
with a batch of shellfish samples collected from Sinclair Inlet. All organic analyses were
conducted by the Department of Ecology/EPA Manchester Laboratory. Metals analyses were
conducted by Columbia Analytical Laboratory. The methods used were standard methods with
the following exceptions:
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Table 2. Analytical methods for Bellingham Bay investigation.
Analysis
Metals
As
Cd
Hg
Pb
Base Neutral Acids
Pest/PCB
Pentachlorophenol
% Moisture
% Lipids
Method
Atomic Absorption 7000
GFAA method 7060
GFAA method 7420
CVAA method 7471
GFAA method 7420
GC/MS method 8270
GC/EC method 8080*
GC/EC method 8150**
Dry @ 105°C
Gravimetric
Reference
EPA 1986a
EPA 1986a
EPA 1986a
EPA 1986a
EPA 1986a
EPA 1986a
EPA 1986a
EPA 1986a
APHA 1985
EPA 1980
Laboratory
Columbia Analytical
Columbia Analytical
Columbia Analytical
Columbia Analytical
Columbia" Analyff&i
Ecology/EPA (Man.)
Ecology/EPA (Man.)
Ecology/EPA (Man.)
Ecology/EPA (Man.)
Ecology/EPA (Man.)
* Chlorpyrifos added to standards and measured in samples.
** Phenoxy herbicide method.
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Table 8. Comparison of metals concentrations in clams with other studies in Puget Sound.
All results ug/g wet weight basis.
Non-Reference Area*
Metal
As
Cd
Pb
Hg
*
**
Elliott Bay
5 sites
Speciei code: 1
Study: Humbert o tl. I9M
Mean
Range
N
Mean
Range
N
Mean
Range
N
Mean
Range
N
Presumption
Used within
2.4
1.8-3.5
?
0.13
.10-. 19
?
0.40
.10-.50
?
0.020
.11-.28
?
Puget Eagle
Sound Harbor
8 sites
2 3
FilpnMum I9M Y«k« ot ). 198«
2.70
1.3-4.1
27
0.32
.10-.54
39
0.09
.04-.18
25
0.02
<.02-.03
25
within study that area
studies as
*** Species codes:
reference or
1
2.9
1.5-4.4
8
0.16
.08-. 29
8
0.84
.43-2.0
8
0.33
McNeil
Island
2
Nonool9M
1.3
1.1-1.4
3
0.31
.29-.34
3
0.011
.01-.07 .010-.011
8
may exceed
control site
3
background
.
2
Reference Area**
Birch Bay Point Horsehead
Blakely Bay
232
FdpnMim I9M Y«ke a .1.1984 Norton 1911
2.6
2.1-3.2
5
- 0.3
.22-.36
6
<.04
<.04-<.04
4
<.02
<.02-<.02
3
concentrations.
3.7 1.4
1 1
0.11 0.35
1 1
0.38
1
0.012 0.012
1 1
3
Bellingham Bay
Post This study
Point 4 sites
2 2
CH2MHUI 1914
1.82
1.1-2.1
4
0.21
.18-. 23
4
0.08
.02-. 15
4
0.01
<.10-.28 <.01-.02
3 4
Saxidomus giganteus Protothaca staminca
Protothaca staminea
Saxidomus giganteus
Tapes japonica
Tresus capax
(first two predominated)
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Table 9. Comparison of PAH concentrations in clams with other studies. All results ug/kg wet weight.
Non-Reference area* Reference Area** Bell. Bay Selected Foods
Chemical
Speciei code""
Study:
LPAH(l)
Mean
Range
N
HPAH(2)
Mean
Range
N
Eagle
Harbor
i
Yikeai). IM4
126
14-690
8
403
45-1575
8
S. Budd Industrial
Inlet Waterway
2 3
Norton I9«« MillM«ill«IO
67 47
16-159 6-96
3 4
373 386
72-938 138-701
3 4
Point
Blakely
i
YttoMtl. 1*14
21
1
76
1
N. Budd
Inlet
2
Norton IM<
<10
1
<10
1
Case This study Smoked Smoked
Inlet 4 sites Ham Fish
3
MtllMol ) 1910 Pucfcmt IH 1 FnckoU 19*1
<1
1
<1
1
3
<2-6
4
3.7
<2-20
4
9.2-145
r
10-496 3-30
Presumption within study that area may exceed background concentrations.
Used within study as reference or control site.
Duwamish, Commencement Bay, and Hylebos Waterways, Seattle Waterfront
Species codes: 1 2
Protothaca stamines
Tapes japonica
Saxidomus giganteus
Protothaca staminca Macoma nasuta
Tapes japonica Macoma carlottensis
Mya arenaria Acila castrensis
(1)
(2)
LPAH - Low molecular weight Polycyclic Aromatic Hydrocarbons. (2 and 3 ring compounds)
HPAH = High molecular weight Polycyclic Aromatic Hydrocarbons. (4,5 and 6 ring compounds)
-------
not found in the samples. DDE was found in low concentrations in a concurrent study in 2 out
of 7 samples and chlordane in 1 out of 7 samples. PAH's were found at low concentrations in
clam samples. Overall, the levels of contaminants examined were low compared with areas with
known sediment contamination and were equivalent to concentrations found at reference areas.
18
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I
REFERENCES CITED
APHA. Standard methods for the examination of water and wastewater. 16th edition. 1985.
CH2M Hill. Final report of Bellingham Bay water quality monitoring program. Prepared for
City of Bellingham and Georgia-Pacific Corporation. CH2M Hill, Bellevue, WA, 1976.
Ecology. Report of findings of the annual wastewater NPDES inspection at Georgia Pacific,
Bellingham, WA. Washington State Department of Ecology, Industrial Section, Olympia,
WA, 1988.
EPA. Manual of analytical methods for the analyses of pesticides in humans and environmental
samples. EPA 600/18-80-038, 1980.
EPA. 1986a. Test methods for evaluating solid waste. EPA Environmental monitoring and
support laboratory, Cincinnati, OH, 1986.
EPA. Recommended guidelines for measuring organic compounds in Puget Sound sediment and
tissue samples. Puget Sound Estuary Program. EPA Region 10, Office of Puget Sound,
Seattle, WA, 1989.
Faigenblum, J. Chemicals and bacteriological organisms in recreational shellfish. Final Report.
Prepared for U.S. Environmental Protection Agency Region 10, Office of Puget Sound,
Seattle, WA, 1988.
FDA. Polychlorinated biphenyls (PCBs) in fish and shellfish; reduction of tolerances; final
decision. Food and Drug Administration. Fed. Reg. V. 49(100); 21514-21520, 1984.
FDA. Action levels for poisonous or deleterious substances in human food and animal feed.
Center for Food Safety and Applied Nutrition, Industry Programs Branch, Food and Drug
Administration. 200 C Street SW, Washington D.C., 1985.
Gahler, A.R., J.M Cummins, J.N. Blazevich, R.H. Rieck, R. Arp, C.E. Gangmark, S.V.W.
Pope, S. Filip. Chemical contaminants in edible, non-salmonid fish and crabs from
Commencement Bay, Washington. USEPA Environmental Services Division, Region X,
Seattle, WA, 1982.
Kendra, W. Investigation of recurrent coho salmon mortality at the Maritime Heritage fish
hatchery in Bellingham WA. Washington State Department of Ecology, Olympia, WA,
1988. 49pp.
19
-------
Malins, D.C., B.B. McLain, D.W. Brown, et dl. Chemical contaminants and biological
abnormalities in Central and Southern Puget Sound. NOAA Technical Memorandum
OMPA-2, 1980. 295 pp.
Norton, D. Results of priority pollutant analyses on water, sediment and clam samples collected
in lower Budd Inlet near McFarland Cascade, Olympia, WA. Washington State
Department of Ecology, Memorandum to Tom Eaton. Olympia, WA, 1986.
Norton, D. McNeil Island: Intertidal screening survey for toxic chemicals in water,sediment
and clam tissue. Washington Department of Ecology, Olympia, WA, 1988.
PSEP. Puget Sound Estuary Program: Recommended protocols for measuring selected
environmental variables in Puget Sound. Final Report. Prepared for U.S. Environmental
Protection Agency Region 10, Office of Puget Sound, 1986.
PSDDA. Management plan report: Unconfined, open-water disposal of dredged material Phase
1 (Central Puget Sound). Puget Sound Dredged Disposal Analysis. U.S. Army Corps of
Engineers, Seattle District; U.S. Environmental Protection Agency, Region 10;
Washington State Department of Natural Resources; Washington State Department of
Ecology, 1988.
Pucknat, A.W. Health impacts of polynuclear aromatic hydrocarbons. Noyes Data
Corporation, Park Ridge, NJ, 1981.
V^'
PTI. Everett Harbor Action Program: Analysis of toxic problem areas. Final Report.
Prepared for U.S. Environmental Protection Agency Region 10, Office of Puget Sound,
Seattle, WA, 1988.
PTI. Bellingham Bay Action Program: Initial data summaries and problem identification.
Prepared for US Environmental Protection Agency Region 10, Office of Puget Sound,
Seattle, WA, 1989.
Rasmussen, L.F., and D.C. Williams. The occurrence and distribution of mercury in marine
organisms in Bellingham Bay. Northwest Science, 49, 87-94. 1975.
Romberg, G.P., S.P. Pavlou, R.F. Shokes, W. Horn, E.A. Crecelius, P. Hamilton, J.T. Gunn,
R.D. Muench and J. Vinelli. Toxicant pretreatment planning study technical report:
Presence, distribution and fate of toxicants in Puget Sound and Lake Washington. Metro,
Seattle, WA, 1984.
SAIC. PSDDA 1990 Crab bioaccumulation survey of Bellingham Bay. Report to Washington
State Department of Natural Resources, Division of Aquatic Lands. Olympia, WA, 1991.
20
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f
Tetra Tech. Pesticides of Concern in the Puget Sound Basin: A review of contemporary
pesticide usage. Final report TC-3338-32 to U.S. Environmental Protection Agency,
1.708.
Yake, B, J. Joy, and A. Johnson. Chemical contaminants in clams and crabs from Eagle
Harbor, Washington State, with emphasis on polynuclear aromatic hydrocarbons.
Washington State J3epartment of Ecology, Olympia, WA, 1984.
21
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APPENDIX 1: LABORATORY QUALITY ASSURANCE
Several tests were used to assess laboratory accuracy and precision. Overall, the data are
usable. Table A-l presents the results of these different tests for organics. Table A-2 shows
results for metals. Following is a review of the tests. These results apply to all samples except
clam site 3B. This sample was collected later and was analyzed along with shellfish from
another study. This sample also passed quality assurance tests.
Matrix Spike: Matrix spikes were performed for each of the three types of analyses. A known
amount of the target compound was added to the matrix (homogenized tissue) and the recovery
of the compound was a measure of extraction efficiency and analytical accuracy. Table 4 shows
all matrix spike recoveries are within acceptable limits.
Replicate Analysis: Relative percent difference (RPD) was calculated from results of replicate
analyses as a measure precision. The formula for RPD is
RPD = (Sl-S2)/((Sl+S2)/2)* 100
where SI and S2 are the duplicate samples. Matrix spike samples were analyzed in duplicate
so that there were two RPD measurements. One sample was split after homogenization and
submitted to the laboratory blind. Results of this blind replicate analysis of metals showed
remarkably similar results, an indication of complete homogenization and high analytical
precision. Because no compounds were found above detection limits in the blind replicate
analyses of organics and pesticides, no blind RPD is available.
Surrogate recovery: For the GC-EC and GC-MS analyses, recovery of surrogates added before
extraction were analyzed. Surrogates have similar chemical structure to the analytes of interest
but are not expected to be found in the environment. The surrogate for PCP analysis is
tribromophenol. For the pesticides, three surrogates, 4,4 dibromooctafluorobiphenyl,
dibutylchlorendate, and octochloronapthalene were used. In the base, neutral and acid
extraction, due to the silica gel cleanup and optimization for PAHs, only the non-polar
surrogates terphenyl-dl4, pyrene-dlO, and 2- fluorobiphenyl were recovered. All surrogates
recoveries were within EPA CLP guidelines for sediment (there are no CLP guidelines for
tissues).
Reference Material: For metals analysis, a standard reference material, oyster tissue, was
analyzed. This material is provided by the National Bureau of Standards and is exhaustively
analyzed and its metals concentrations certified to be within a narrow range of values. Results
showed high accuracy.
Method Blanks: Analysis of method blanks showed no laboratory contamination.
22
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t
t
Table A-l. Results of matrix
spike
recovery tests for organics.
Percent Spike Recovery
Crab
Lab number
PESTICIDES
Aldrin
Chlordane
4,4'-DDT
alpha Endosulfan
Endrin
Heptachlor
gamma-BHC
Methoxychlor
Pentachlorophenol
PAHs
Napthalene
Acenaphthylene
Acenapthene -
Fluorene
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benzo(a)Anthracene
Chrysene
Benzo(b)Fluoranthene
Benzo(k)Fluoranthene
Benzo(a)Pyrene
Ideno(l,2,3-cd)Pyrene
Dibenz(a,h)Anthracene
Benzo(g,h,i)Perylene
8081
101%
85%
81%
84%
82%
76%
75%
95%
99%
--
~
8090 RPD(l)
98%
81%
71%
84%
82%
72%
80%
94%
105%
"
3%
5%
14%
0%
1%
5%
6%
1%
6%
--
*
~
"
Clam
8093
108%
122%
96%
100%
103%
88%
95%
116%
46%
73%
78%
91%
83%
70%
74%
120%
103%
102%
88%
96%
91%
85%
73%
75%
8093 RPD
96%
102%
94%
99%
94%
88%
94%
129%
66%
85%
83%
96%
86%
73%
76%
138%
114%
111%
104%
98%
96%
82%
75%
51%
12%
18%
1%
1%
9%
0%
1%
11%
36%
15%
6%
5%
4%
4%
3%
14%
10%
8%
17%
2%
5%
4%
3%
38%
Recommended
Range for spike(2)
50%-150%
50%-150%
50%-150%
50%-150%
50%-150%
50%-150%
50%-150%
50% -150%
50%-150%
50%-150%
50%-150%
50% -150%
50%-150%
50%-150%
50%-150%
50%-150%
50%-150%
50%-150%
50%-150%
50% -150%
50%-150%
50%-150%
50%-150%
50%-150%
(1) Relative Percent Difference
(2) From Puget Sound Estuary Program Guidelines (EPA 1989). Due to problems of matrix
interference, exceedence of spike recovery limits does not require data qualification.
23
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Table A-2. Results of tests of laboratory accuracy and precision for metals.
Spike Recovery
Lab no.
Arsenic
Cadmium
Lead
Mercury
Crab
8087
58%
94%
103%
100%
Clam
8094
100%
89%
94%
100%
8094
14.7
1.53
0.4
0.1
Precision of multiple analyses (results=mg/kg)
Lab
8094
14.1
1.62
0.4
0.1
Blind(2)
RPD(l)
4%
6%
<1%
<1%
8087
21.8
<0.01
<0.2
0.8
8091
22
0.01
<0.2
0.8
RPD
1%
NA
NA
0%
8093 8097
13.2 11.9
1.86 1.84
1.2 1.1
<0.1 <0.1
RPD
10%
1%
9%
NA
Standard Reference
Material (3)
True Found %
14.0 13.8 99%
4.15 4.1 98%
0.371 0.40 108%
0.064 0.06 93%
(1) Relative Percent Difference
(2) Homogenized split of sample submitted to laboratory as separate sample
(3) National Bureau of Standards oyster tissue #1566a
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MANCHESTER ENVIRONMENTAL LABORATORY
7411 Beach Drive SE , Port Orchard Washington 98366
CASE NARRATIVE
December 13,1990
Subject: Bellingham Bay Bioaccumulation
Samples: 90 - 398080, 398081, 398082, 398083, 398085, 398086, 398087, 398090, 398091, 398093,
398094, 398096, 398097
Case No. DOE-601B
Officer: James Cubbage
By: Dickey D. Huntamer
Robert Carrell &S
Organic Analysis Unit
FOLHVtfCIEAK AROMATIC HYDROCARBONS
ANALYTICAL METHODS:
No official EPA method exists for semivolatile tissue analysis. The prepared tissue samples were
extracted with a 50:50 mixture of methylene chloride and acetone using the Manchester modification of
the EPA CLP and SW-846 Method 8270 procedure with capillary GC/MS analysis of the sample
extracts. All CLP QA/QC procedures were performed on the samples. Since Polynuclear Aromatic
Hydrocarbons (PAH) were the primary target analytes and low detection limits were desired sample
cleanup using Gel Permeation Chromatography (GPC) at both 2000 Molecular Weight (MW) and
1000MW cutoff (SW-846 Method 3640) followed by Silica Gel cleanup Method 3630 was done on the
samples. Lower Quantitation Limits were also realized by extracting approximately 50 grams of tissue
and concentrating the final extract to 1.0 mL for analysis.
HOLDING TIMES:
Under Puget Sound Estuary Program (PSEP) Guidelines for organic compounds tissue samples can be
stored frozen for up to one year before extraction. After collection samples were prepared for the
laboratory by the field staff and stored frozen until extraction. Since the samples were stored frozen all
sample extraction holding times were met. The reporting form for holding times indicates that the
sample holding times were exceeded however this is not he case since it is measured from collection
date and includes the time the samples were frozen. All analyses were performed within the specified
40 day holding time.
BLANKS:
No significant PAH blank contamination was detected.
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Bellingham Bioaccumulation - Tissue
SURROGATES:
The samples received all six surrogate compounds normally added to semivolatile analyses. Due to the
silica gel cleanup only the non-polar surrogates Terphenyl-dl4, Pyrene-dlO and 2-Fluorobiphenyl were
recovered. Only one of these compounds, Pyrene-dlO is a true PAH compound and is representative
of the PAH target analytes. Surrogate spike recoveries for all three compounds were within normal
limits for CLP soil recovery limits. The CLP recovery limits are only advisory since no tissue surrogate
spike recovery limits have been established.
MATRIX SPIKE AND MATRIX SPIKE DUPLICATE:
Matrix spikes compounds were added at 20 ug, rather than the normal spiking concentration of 50 ug,
to more closely approximate the low detection limits requested. No significant problems were
encountered with recovering the matrix spike compounds at this level (400 ug/Kg wet weight).
Although no matrix spike recovery limits have been established at this low level, spike recoveries were
generally within the normal CLP recovery range found at higher matrix spike levels.
Two matrix spike and matrix spike duplicates (MS/MSD) were analyzed with the set. Sample 398093
was used as one matrix source and due to insufficient sample tissue from both 398081 and 398090 had
to be used to make the second MS/MSD pair. Matrix spike recoveries ranged from 48% to 103 % for
398081/398090 and 46% to 138% for 398093.
SPECIAL ANALYTICAL PROBLEMS:
No analytical problems were encountered in the analysis. The low detection limits were achieved by
extracting SO grams of sample and concentrating the extract after cleanup to 1.0 mL prior to analysis.
PESTICIDES IPCB - CHLORPYRIFOS AND PCP
ANALYTICAL METHODS:
The tissue (clams and crabs) was extracted by the Manchester Laboratory using a Polytron tissue
grinder and a 50:50 mixture of methylene chloride and acetone as the solvent. The analyses were done
following EPA Method 8080 (chlorinated pesticides, PCB's and chlorpyrifos) and EPA Method 8150
(PCP) using capillary Gas Chromatography/Electron Capture Detector (GC/ECD) analysis.
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Bellingham Bioaccumulation - Tissue j
The percent lipid determination was performed in a similar fashion to the analytical extractions except
petroleum ether was used as the solvent. The extract was evaporated and weighed to determine the
extractable lipid. Percent solids were also determined on the samples. The results are given in the
table below.
Lab Number Percent Solids Percent Lipids
398080 16 0.0
398081 16 0.09
398082 16 0.0
398083 17 0.09
398085 16 0.0
398086 15 0.0
398087 19 0.0
398090 13 0.10
398091 19 0.09
398093 11 0.30
398094 14 0.39
398096 14 0.68
398097 11 0.10
BLANKS:
No significant blank contamination was found.
HOLDING TIMES:
All samples were analyzed within the 40 day holding time.
SURROGATES:
Surrogate spike recoveries for the Pesticides/PCB's ranged from 70.1 % to 117% for Dibromoocta-
fluorobiphenyl (DBOB); 73.3% to 108.8% for Dibutylchlorendate (DEC); 63.7% to 118.3% for
octachloronaphthalene (OCN) and 71.9% to 115.3% for Decachlorobiphenyl (DCB). These values are
well within the advisory limits of 60% to 150% recoveries listed in CLP for soil samples.
For the PCP analysis, excluding the method blanks which experienced low recoveries due to lack of
"keeper", the surrogate recoveries for Tribromophenol (TBP) ranged from 75.1 % to 99.6%. The
method blank surrogate recoveries were 26.7% to 86.1 %. There are, however, no advisory lim;
data qualifiers were not added to the data based on surrogate recoveries.
MATRIX SPIKE AND MATRIX SPIKE DUPLICATE:
Four matrix spikes were analyzed for pesticides to reflect the two different types of tissue
(clam and crab) matrix effects. Recoveries of the pesticides ranged from 70.5% to 129.3%.
Two matrix spikes were run for PCP with recoveries of PCP ranging from 99.1 % to 104.9%.
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Bellingham Bioaccumulation - Tissue
SPECIAL ANALYTICAL PROBLEMS:
The pesticides were run on the tissue extracts first then the extract was cleaned up with Sulfuric acid
treatment. These acid treated extracts were then reanalyzed for PCB's thus allowing lower quantitation
limits.
DATA QUALIFIER CODES:
U - The material was analyzed for, but was not detected. The associated
numerical value is the sample quantitation limit.
J - The associated numerical value is an estimated quantity.
R - The data are unusable (compound may or may not be present). Resampling
and reanalysis is necessary for verification.
NAR No Analytical Result.
M - The compound was detected and confirmed but was not quantitated.
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