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Microchemical Journal 88 (2008) 52 – 55

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Determination and comparison of heavy metals in selected seafood, water,

vegetation and sediments by inductively coupled plasma-optical emission
spectrometry from an industrialized and pristine waterway
in Southwest Louisiana
Michael A. Hamilton, Paul W. Rode, Mark E. Merchant, Joseph Sneddon ⁎
Department of Chemistry, McNeese State University, Lake Charles, Louisiana 70609, USA
Received 19 September 2007; accepted 27 September 2007
Available online 6 October 2007

Abstract

Samples of crab, fish, sediments, vegetation and waters were collected from a pristine (Rockefeller Wildlife Refuge) and historically polluted
(Bayou d'Inde) waterway in Southwest Louisiana. After sample preparation via microwave digestion to obtain a solution, cadmium, mercury,
nickel and lead were determined by inductively coupled plasma-optical emission spectrometry. A comparison of results showed that there was
very little difference in concentrations for the four metals between the two areas, typically in the 10 μg/g range for crabs and fish, around 0.05 μg/
mL or lower for waters, and somewhat higher for sediments and vegetation of around two to three times.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Metals; ICP-OES; Crabs; Vegetation; Fish; Sediments; Waters

1. Introduction documented and historically polluted waterway and the

Southwest Louisiana is one of the most productive areas in the
continental USA for fishing, both commercial and recreational.
Because of its remoteness and nearness to water in the 1940's it
was chosen as an area for fossil fuel refining and production of
associated petrochemicals. None or poorly enforced environmen-
tal laws resulted in widespread pollution, particularly from heavy
metals such as cadmium, mercury and lead. While the intro-
duction and enforcement of environmental laws has significantly
improved from the early 1970's, the possibility of heavy metal
contamination and sources of previously polluted waterways still
exist in Southwest Louisiana [1].
The object of this study was to compare the concentration
of selected heavy metals (cadmium, mercury, nickel, and
lead) in two quite different waterways: one on a well
⁎ Corresponding author.
E-mail address: jsneddon@mcneese.edu (J. Sneddon).
0026-265X/$ - see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.microc.2007.09.004
second a more pristine waterway. Of particular interest was
the concentration of these metals in crabs, fish, vegetation,
sediments and waters. Crabs and fish are frequently caught
and consumed by recreational fisherman, often a family
tradition [2].
2. Experimental
2.1. Sampling sites and collection
The two areas investigated are on the heavily industrialized
and historically polluted Bayou d'Inde waterway, and the more
pristine Rockefeller Wildlife Refuge, both located in Southwest
Louisiana.
Bayou d'Inde is located approximately one mile west of the
city of Lake Charles, Louisiana. The waterway empties into
Lake Charles. Bayou d'Inde is located in the northern part of the
Calcasieu Estuary, west of the city of Lake Charles in Calcasieu
Parish, Louisiana [1,2]. Bayou d'Inde's headwaters originate in
 M.A. Hamilton et al. / Microchemical Journal 88 (2008) 52–55
Table 1
Sampling location on Bayou d'Inde
Site GPS (coordinates) Samples taken
1 N 30° 12.289′ Water, vegetation, sediment, crab, fish
W 93° 20.566′
2 N 30° 12.102′ Water, vegetation, sediment, crab, fish
W 93° 19.7771′
3 N 30° 12.129′ Water, vegetation
W 93° 19.490′
4 N 30° 12.720′ Water, vegetation
W 93° 18.632′
5 N 30° 12.429′ Water, vegetation
W 93° 18.368′
6 N 30° 12.480′ Water, vegetation
W 93° 17.841′
the western part of Sulphur, Louisiana and flow primarily east-
southeast through heavy commercial and industrial areas
emptying into the Calcasieu Ship channel west of Lake Charles.
Bayou d'Inde is a tidally influenced wetland bayou that
flows through or adjacent to property owned by Citgo
Petroleum (Citgo), OxyChem Corporation (OxyChem), Fire-
stone Synthetic Rubber and Latex Company (Firestone),
Westlake Polymers Corporation (Westlake Polymers), Brown-
ing-Ferris Industries (BFI), Certain-Teed Products Corporation
(Certain-Teed), and Pittsburgh Paint and Glass Industries
(PPG). These facilities are all active.
The land around Bayou d'Inde includes undeveloped
wooded marshland, rural residential, commercial, and heavy
industrial property. Rural residential and underdeveloped
woodland areas border the Bayou northwest and up gradient
of the industrial area. Heavy industry dominates the middle and
southern reaches of Bayou d'Inde on both sides. Several of the
industries have wastewater outfalls permitted under the
National Pollutant Discharge Elimination System (NPDES).
Permitted discharges to Bayou d'Inde include outfalls belong-
ing to Citgo, OxyChem, Firestone, Westlake Polymers, BFI,
Certain-Teed, and PPG.
In addition, PPG discharges wastewater to Bayou d'Inde via
the PPG Canal, which enters the bayou approximately one mile
upstream of the Calcasieu Ship Channel. These discharges
(current and historic), storm-water runoff, and accidental re-
leases have contributed to organic and inorganic impacts to
surface water, sediment, and biota within the Bayou d'Inde area
of concern.
Although Bayou d'Inde is not used as a drinking water
source, the estuary surface waters have been designated by the
Louisiana Department of Environmental Quality as supporting
primary contact recreation, secondary contact recreation, and
fish and wildlife propagation. Bayou d'Inde supports recrea-
tional fishing and has several delineated wetlands that are
considered sensitive environments. Health advisories warning
of contaminated fish consumption have been issued for the
Calcasieu Estuary including Bayou d'Inde.
The Bayou d'Inde area of concern has not been proposed for
inclusion on the National Priorities List (NPL), but the entire
Calcasieu Estuary has been the subject of environmental studies
dating back to the early 1970's.
53
The sampling of this project involved six locations
throughout the Bayou D'Inde area. A global positioning system
(GPS) was used to establish the exact location of each site and
shown in Table 1. The sampled substances to be tested included:
(1) Fish (mullet), (2) Crab (Blue Crab, Callinectes sapidus),
(3) Vegetation, (4) Water, and (5) Sediments. The samples were
collected on two different dates, 01/26/06, temperature of 20 °C,
samples 51–79 (water, sediments and vegetation) and 05/30/06,
temperature of 33 °C, samples 1–50 (samples 42–50 from
sampling Site 1 of whole crab were contaminated and results not
presented in this work).
The water samples were collected by scooping a plastic
container into the water in the sampling area in a 50 mL plastic
container. The soil was collecting by using a four foot PVC pipe
about 5 ft from the shoreline. The pipes were labeled and stored
in cold storage room (0 °C). The vegetation was collected from
the shoreline using a catch and grab method. Flora was stored in
Ziploc bags, labeled and stored at − 20 °C. The crabs and fish
were collected using a hybrid fish/crab trap. Fish and crabs were
individually stored in Ziploc bags, labeled and stored at − 20 °C.
The crabs collected were only found in Site 1 and 2. The other
sites did not produce crabs. The fish collected were only found
in Site 1 and 2.
Rockefeller Wildlife Refuge is a federally protected refuge,
and harbors various types of wildlife. It is located in eastern
Cameron and western Vermilion Parishes, South of Lake
Charles (approximately 40 miles) and is owned and maintained
by the State of Louisiana. This area, of approximately 76,000 ac
borders the Gulf of Mexico for 26.5 miles and extends inland
toward the Grand Chenier ridge, a stranded beach ridge, and
6 miles from the Gulf. It is remote from industry and is regarded
as a pristine area with various wildlife studies such as alligator
research in progress. Detailed information about Rockefeller
Refuge can be found elsewhere [3]. The samples were acquired
from the Wildlife Employees located at Rockefeller Refuge
testing complex. The whereabouts of these samples are not
exact. Samples are known to be caught near the middle of the
refuge. The samples were acquired on 08/03/06. Acquisitions
included blue crab and fish. Samples acquired were labeled 80–
117.
2.2. Sample preparation
Sample preparation was similar to that described previously
[4]. A few drops of concentrated nitric acid was added to all
water samples and stored in at 4 °C prior to determination by
inductively coupled plasma-optical emission spectrometry
(ICP-OES). The vegetation was removed from cold storage
and placed in a mortar and dried overnight at 70 °C in an oven.
The sample was ground with the pestle and approximately
0.3000 g accurately weighed into the microwave digestion tube,
and 4 mL of concentrated nitric acid and 2 mL of hydrogen
peroxide then added. The sample was microwaved on power 3
for 3 min, and removed and placed under a hood for 5 min. A
microwave of a further 5 min at microwave power of 3 was
followed by a 5 minute cool down in a hood. The microwave
digestion was vented for 5 min. The contents were filtered into a
54 M.A. Hamilton et al. / Microchemical Journal 88 (2008) 52–55

25.0 mL or 50.0 mL volumetric flask and diluted with de-
ionized water. The sample was stored in a plastic container at
4 °C prior to determination by ICP-OES.
Sediments were collected as previously described [4] and
involved using a four foot PVC pipe about 5 ft from the shoreline.
The pipes were labeled and stored at 0 °C. Prior to metal
determination they were dried overnight at 70 °C in an oven. The
sample was ground with a pestle and approximately 0.3000 g
accurately weighed into the microwave digestion tube and
identical digestion as the vegetation performed.
The blue crab was removed from cold storage and either
dissected or left whole. The dissected crab was opened and the
gills, meat and shell dried at 70 °C overnight. Following
grinding in a mortar and pestle four samples of accurately
weighed approximately 0.3000 g was prepared as the vegetation.
Fish was prepared in an identical fashion to that of the blue
crab.
conditions are described in detail elsewhere [5]. Results were
determined in μg/mL (parts per million) and converted into μg/g
(except waters) as follows:
lg=mL
50:0 mL or 25:0 mL
: ð1Þ
accurate mass of approximately 0:3000 g of sample
Detection limits (signal-to-noise ratio of three) for solution
determination of the four metals were around 0.02 μg/mL which
translates into detection limits of around 1.67 μg/g for solid
samples using Eq. (1) with a 25.0 mL volume.
2.4. Chemicals
All chemicals were reagent grade. Stock solutions of
1000 μg/mL (ppm) were used to prepare calibration standards
daily as required.

2.3. Instrumentation 2.5. Accuracy

All quantitative metal determination was performed using a Recoveries were periodically run during the sample pre-
Leeman Labs PS Series (Leeman Labs, Lowell, Massachusetts) paration and subsequent determination for all four metals
inductively coupled plasma-optical emission spectrometer resulting in values in an acceptable range of 92–95%. National
(ICP-OES). The instrument setting, wavelengths and operating Institutes of Standards & Technology (NIST) (Gaithersberg,

Table 2
Concentrations of cadmium, nickel, lead and mercury in crabs, fish, vegetation, sediments (in μg/g), and waters (μg/mL) from Bayou d'Inde and Rockefeller Wildlife
Refuge
Cd Ni Hg Pb
Bayou d'Inde
A. Sampling 1
1. Whole crabs, 1–12, Site 1 8.17 ± 0.11 (8.00–8.36) 8.09 ± 0.50 (7.44–9.31) 12.16 ± 0.86 (11.12–14.07) 4.39 ± 2.04 (1.70–6.53)
2. Whole crabs, 36–41, Site 2 4.17 ± 0.06 (4.08–4.24) 6.90 ± 1.47 (6.42–9.04) 5.91 ± 0.37 (5.84–6.46) 2.62 ± 2.09 (1.80–6.16)
3. Crab dissections, 22–28, Site 1 4.52 ± 0.89 (4.02–6.34) 12.61 ± 6.61 (6.73–24.67) 6.30 ± 0.95 (5.52–8.63) 3.46 ± 2.56 (2.00–7.91)
4. Crab dissections, 29–35, Site 2 4.20 ± 0.23 (4.07–4.15) 7.73 ± 4.94 (4.86–18.66) 6.31 ± 0.41 (5.90–7.08) 4.61 ± 2.66 (1.67–9.21)
5. Whole fish 13–21, Site 2 9.05 ± 1.67 (7.72–11.58) 9.65 ± 2.94 (5.95–14.45) 13.42 ± 2.41 (11.41–17.03) 3.68 ± 2.74 (1.70–6.16)

B. Sampling 2
6.a Vegetation 63–64, Site 1 4.46 (4.45, 4.46) 12.80 (12.48, 13.21) 6.41 (6.56, 6.25) 15.02 (14.44, 15.60)
6.b Vegetation 65–66, Site 2 4.58 (4.64, 4.51) 14.54 (12.92, 16.15) 6.69 (6.76, 6.61) 19.40 (28.12, 10.67)
6.c Vegetation 67–68, Site 3 4.26 (4.24, 4.29) 14.91 (10.28, 19.53) 6.36 (6.42, 6.30) 11.19 (10.28, 12.10)
6.d Vegetation 69–70, Site 4 4.36 (4.49, 4.22) 14.61 (17.95, 11.27) 6.25 (6.58, 5.91) 11.75 (12.10, 9.40)
6.e Vegetation 71–72, Site 5 4.77 (4.97, 4.61) 18.43 (20.79, 16.07) 6.25 (6.19, 6.32) 25.03 (28.99, 21.09)
6.f Vegetation 73–74, Site 6 4.55 (4.57, 4.53) 16.37 (16.87, 15.86) 6.14 (6.27, 5.99) 16.87 (17.44, 16.26)
6.g Sediments 75, 78, Site 1 4.40 (4.46, 4.33) 18.33 (12.60, 24.05) 6.20 (6.03, 6.37) 8.33 (6.96, 9.69)
6.h Sediments 76, 79, Site 2 (77–none) 4.38 (4.50, 4.26) 16.75 (14.99, 18.50) 6.22 (6.00, 6.43) 7.60 (6.97, 8.23)
7.a Water 51–52, Site 1 0.05 (0.05, 0.05) 0.06 (0.06, 0.06) 0.08 (0.08, 0.08) 0.03 (0.03, ⁎)
7.b Water 53–54, Site 2 0.05 (0.05, 0.05) 0.05 (0.05, 0.05) 0.07(0.07, 0.07) ⁎
⁎
7.c Water 55–56, Site 3 0.05 (0.05, 0.05) 0.06 (0.05, 0.06) 0.07 (0.07, 0.07) ⁎
⁎
7.d Water 57–58, Site 4 0.05 (0.05, 0.05) 0.06 (0.06, 0.05) 0.08 (0.08, 0.07) 0.03 (⁎, 0.03)
7.e Water 59–60, Site 5 0.05 (0.05, 0.05) 0.06 (0.05, 0.06) 0.06 (0.06, 0.06) ⁎ (⁎)
7.f Water 61–62, Site 6 0.05 (0.05, 0.05) 0.05 (0.05, 0.05) 0.08 (0.07, 0.08) 0.04 (0.04, ⁎)

b. Rockefeller Refuge
A. Sampling 2
1.Whole crabs 80–95 8.31 ± 0.36 (8.08–9.60) 8.58 ± 1.29 (6.32–10.68) 11.49 ± 0.39 (10.37–12.07) 6.76 ± 3.26 (2.42–11.06)
2. Crab dissections, 96–101A, 101B, 106A 4.09 ± 0.22 (3.83–4.65) 4.27 ± 2.73 (2.54–11.61) 5.56 ± 0.29 (5.47–6.02) 3.29 ± 2.14 (1.77–6.56)
3. Whole fish 106B–117 8.19 ± 0.16 (7.96–8.58) 8.14 ± 1.04 (6.58–10.22) 11.44 ± 0.32 (10.39–12.36) 7.89 ± 3.43 (1.77–12.12)
⁎Less than detection limit.
 M.A. Hamilton et al. / Microchemical Journal 88 (2008) 52–55
Maryland) — Standard Reference Material, SRM-1577b bovine
liver was used. Results for cadmium and lead were statistically
acceptable to the standard. The concentration of the NIST-SRM-
1577b for mercury is 0.004 μg/g and was not detected by the ICP-
OES instrumentation. There was no certified value for nickel.
Quality assurance/quality control (QA/QC) was used as described
previously [4].
3. Results and discussion
The results of this study are summarized in Table 2. Each
sample (numbers 1–117, except 42–50) was determined three
times with a relative standard deviation (RSD) of better than
0.5% in all cases (results not shown). The results of each
sampling sub-area, i.e., whole crabs, etc. were pooled with the
mean, ± standard deviation and range presented for each pooled
sample.
In general, there was no statistical difference in concentra-
tions for the four metals between the two sampling sites for
whole crab, crab dissections or fish. In general, concentrations
for cadmium, mercury and nickel were around 10 μg/g or lower
and for lead were about a factor of two lower than the other
three metals. For vegetation the concentrations of lead and
nickel were significantly higher than that of cadmium and
mercury. In sediments the concentrations of all four metals was
comparable and below 10 μg/g. In waters the concentrations of
all four metals was in the very low sub-ppm range.
4. Conclusion
Results from this field study show that there are small
differences (in most cases less than a factor of 2) for the metals
(cadmium, lead, nickel, and mercury) determined in crabs, fish,
vegetation, waters and sediments in this study between the
potentially polluted waterway (Bayou d'Inde) and the more
pristine waterway (Rockefeller Wildlife Refuge). It is possible
that the historically polluted waterway has less pollution (in
terms of these four metals) at present than has been found in
55
studies dating back to the 1970's and earlier. Alternatively it is
possible that Rockefeller Wildlife Refuge is not immune or
isolated from metal pollution. A potential factor in this area was
the presence of Hurricane Rita in late September 2005 which
affected much of Southwest Louisiana and in particular
Cameron Parish which could upset the fragile balance between
pristine and more industrialized areas.
This field study provides useful information and a baseline
for future and continued studies on the metal concentrations
in crabs, fish, vegetation, waters, and sediments in Southwest
Louisiana. The work is a continuation of previous studies [6].
Acknowledgements
This work was supported, in part, by Merck undergraduate
research program awarded to McNeese State University for
2005–2007 and provided stipends for two of the authors (MAH
and PWR). Partial support from Environmental Protection
Agency, EPA-R-82958401-1 is gratefully acknowledged.
References
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[3] http://www.wlf.louisiana.gov/experience/wmas/refuges/rockefeller.cfm.
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