Food

Chemistry
Food Chemistry 100 (2007) 853858
www.elsevier.com/locate/foodchem

Analytical, Nutritional and Clinical Methods

Determination of mercury in sh by cold vapour atomic

absorption spectrometry using an automatic mercury analyzer
a,* b
R.B. Voegborlo , H. Akagi
a
Chemistry Department, Kwame Nkrumah University of Science and Technology, KNUST Campus, Kumasi, Ghana
b
National Institute for Minamata Disease, Minamata, Kumamoto 867, Japan

Received 3 August 2004; received in revised form 30 August 2005; accepted 25 September 2005

Abstract

The concentration of mercury in sh samples from the Atlantic coast of Ghana was determined using a simple, rapid and accurate
method. A mixture of HNO3, HClO4 and H2SO4 was used for complete oxidation of organic tissue. Mercury is detected by the cold
vapour atomic absorption spectrometry technique using an automatic mercury analyzer developed at the National Institute for Mina-
mata Disease (NIMD). In total, 56 samples covering 13 species of sh were analysed for total mercury. The concentration of mercury in
the edible muscle tissue of these sh ranged from 0.004 to 0.122 lg g 1 wet weight. All sh species sampled had concentrations less than
the WHO limit of 0.5 lg g 1 wet weight. The low concentrations of mercury detected in the samples do not constitute any signicant
mercury exposure to the general population through sh consumption.
2005 Elsevier Ltd. All rights reserved.

Keywords: Fish; Mercury; Cold vapour atomic absorption spectrometry; Automatic mercury analyzer

1. Introduction in sh mercury concentrations (Hakanson, Nilson, &

Mercury contamination of the marine environment has
long been recognized as a serious environmental concern.
It is widely recognized that human activities have arti-
cially increased mercury loads in the atmosphere on a local,
regional and even hemispheric scale, leading to the contam-
ination of the environment (Slemr & Langer, 1992; Thomp-
son, Furnes, & Walsh, 1992). Population growth and
urbanization have contributed to signicantly elevated lev-
els of mercury in the atmosphere and it has been estimated
that mercury derived from anthropogenic activities in the
atmosphere is up to 80% of the total mercury in the atmo-
sphere (Mason, Fitzgerald, & Morel, 1994). The enhanced
atmospheric deposition of mercury is often the dominant
source of mercury to the aquatic systems, which may reect
*
Corresponding author. Tel.: +233 24 4234318; fax: +233 51 60305.
E-mail addresses: raybrightv@yahoo.com, rbvoegborlo.sci@knust.
edu.gh (R.B. Voegborlo).
Andersson, 1988; Rolfhus & Fitzgerald, 1995).
Since the tragedy of Minamata Bay in Japan (Kurland,
Faro, & Seidler, 1960) most concern has centred on the
presence of mercury in sh since seafood is a major source
of this element. With the exception of occupational expo-
sure, sh are acknowledged to be the single largest source
of mercury to man. Fish accumulate substantial concentra-
tions of mercury in their tissues and thus can represent a
major source of this element to humans. This has been a
matter of concern since its toxicity was clearly documented
(Uchida, Hirakawa, & Inoue, 1961). Mercury, particularly
in the form of methylmercury, is extremely toxic to marine
organisms, wildlife, and man. The main pathway for hu-
man exposure to methylmercury is through consumption
of shery products. The likelihood of mercury toxicity
from sh consumption has been identied in Peru and
some coastal regions of the Mediterranean (Inskip & Pio-
trowski, 1985; Piotrowski & Inskip, 1981). In some in-
stances, sh catches have been banned for human
consumption because their total mercury content exceeded

0308-8146/$ - see front matter 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2005.09.025
854 R.B. Voegborlo, H. Akagi / Food Chemistry 100 (2007) 853858
the maximum limits recommended by the Food and Agri-
culture/World Health Organisation (FAO/WHO, 1972).
Consequently, extensive surveys have been carried out in
a number of countries to evaluate the presence of mercury
in the aquatic biota including sh. Mercury also biomagni-
es through the food chain; so large predatory sh species
tend to have higher levels than non-predatory sh species
at lower levels in the food chain. The establishment of max-
imum permissible mercury concentrations in sh for hu-
man consumption in the range of 0.51.0 lg g 1 wet
weight by many countries has triggered a process of survey-
ing mercury concentrations in natural sh populations
(Lacerda et al., 2000). Recently, levels of mercury in sh
have been widely reported (Lacerda et al., 2000; Lasorsa
& Gill, 1995; Love, Rush, & McGrath, 2003; Monteiro,
Costa, Furness, & Santos, 1996; Nakagawa, Yumita, &
Hiromoto, 1997; Nixon, Rowe, & McLaughlin, 1994; Rolf-
hus & Fitzgerald, 1995; Storelli, Giacominelli-Stuer, &
Marcotrigiano, 2002; Storelli, Stuer, Storelli, & Marcot-
rigiano, 2003 Voegborlo, El-Methnani, & Abedin, 1999;
WHO, 1976). However, information on mercury levels in
marine organisms from the African Coast region is
unavailable. Consequently, no work has been undertaken
in Africa to study human exposure to mercury through
the consumption of sh.
Due to the lack of any comprehensive data on the Hg
content of sh from this part of the Atlantic Ocean and
the considerable global concern about mercury contamina-
tion of commercial and recreational shery products, a sur-
vey of Hg concentrations in dierent species of sh from the
coastal waters of Ghana has been initiated at the Chemistry
Department of the Kwame Nkrumah University of Science
and Technology, in order to determine whether mercury
occurs in marine sh from the coastal waters of Ghana at
concentrations of potential human health concern. The lev-
els will also provide the basis for assessing long-term trends.
This survey is expected to involve the analysis of several dif-
ferent species of marine sh representing dierent trophic
levels in the marine food chain. There is therefore the need
to use rapid and reliable techniques, requiring minimum
analysis time and suited for the routine analysis of large
numbers of sh samples. This paper reports result of Hg
concentrations in a variety of species from the coastal
waters of Ghana obtained using a procedure, which was
developed at the National Institute for Minamata disease
in Japan (NIMD) by Akagi and Nishimura (1991) with
slight modications. It is hoped that the results of this study
will aid in generating data needed for the assessment of mer-
cury intake from sh. Such data is needed for the develop-
ment of consumption advisories for the general public.
2. Materials and methods
2.1. Apparatus
All glassware used were soaked in detergent solution
overnight; rinsed and soaked in 10% (v/v) HNO3 over-
night. They were rinsed with distilled water followed by
0.5% (w/v) KMnO4 and nally rinsed with distilled water
before use.
Automatic Mercury Analyzer Model HG-5000 (Sanso
Seisakusho Co., Ltd., Japan), equipped with mercury lamp
operated at a wavelength of 253.7 nm was used for the
determinations. The signals were obtained on a Yokogawa
Model 3021 strip chart recorder.
Digestion apparatus was thick walled long neck 50 ml
volumetric asks and a block heater with a temperature
range of 150350 C.
2.2. Reagents
All reagents used were of analytical reagent grade (BDH
Chemicals Ltd., Poole, England) unless otherwise stated.
Double distilled water was used for the preparation of all
solutions.
Mercury stock standard solution (1000 mg L 1) was
prepared by dissolving 0.0677 g of HgCl2 in the acid mix-
ture HNO3H2SO4HClO3 (2 + 10 + 2) in a 50 ml diges-
tion ask with heating on a hot plate at a temperature
between 150 and 250 C until the solution became clear.
The solution was then diluted to 50 ml with water. Blank
solutions were also prepared alongside and bulked together
for use as a diluent. The working solutions were freshly
prepared by diluting an appropriate aliquot of the stock
solution through intermediate solutions using blank solu-
tion. Stannous chloride solution (10% w/v) was prepared
by dissolving 10 g of the salt in 100 ml of 1 M HCl. The
solution was aerated with nitrogen gas at 50 ml min 1 for
30 min to expel any elemental mercury from it.
2.3. Sampling and sample preparation
The sh species were collected from random commercial
catches landed at a local shing port in James Town, Accra
between November 2003 and January 2004 in three
batches, depending on the species available for sale. Sam-
ples obtained were therefore reective of species meant
for consumption. A total of fty-six (56) samples, covering
thirteen (13) dierent species were obtained. The samples
were sorted by species, placed in clean plastic bags and
stored on ice in an ice chest. They were then transported
to the laboratory, identied and kept in a freezer at
20 C prior to preparation for chemical analysis. The
samples were washed with distilled water and dried in tis-
sue paper after defrosting in the laboratory. A portion of
the edible muscle tissue was removed from the dorsal part
of each sh, homogenized and stored in clean-capped glass
vials and kept in a freezer until analysis.
2.4. Digestion procedure
The sh samples were digested for total mercury deter-
mination by an open ask procedure developed at the Na-
tional Institute for Minamata Disease (NIMD) in Japan by
 R.B. Voegborlo, H. Akagi / Food Chemistry 100 (2007) 853858
Akagi and Nishimura (1991). The accuracy of this method
has been veried at NIMD through interlaboratory com-
parison exercises (Malm et al., 1995) and by participating
in the analyses of Certied Reference Materials (CRMs)
(e.g. IAEA 085, 086 and 142) supplied by the International
Atomic Energy Agency (IAEA). In the procedure, 0.5 g of
homogenized sh sample was weighed into 50 ml volumet-
ric digestion ask and a mixture of 1 ml H2O, 2 ml HNO3
HClO3 (1:1) and 5 ml H2SO4 was added. The mixture was
then heated at a temperature between 150 and 250 C until
the solution was clear. The sample solution was then
cooled and diluted to 50 ml with double distilled water. A
blank and standard solution digests using 25, 50 and
100 ll of 1 lg/ml standard Hg solution were subjected to
the same treatment. The concentrations of the standard
solution digests obtained were 0.5, 1.0 and 2.0 ng/ml.
2.5. Determination of mercury
Determination of mercury in all the digests was carried
out by cold vapour atomic absorption spectrophotometry
using an Automatic Mercury Analyzer Model HG-5000
(Sanso Seisakusho Co., Ltd., Japan) developed at NIMD.
The analyzer consists of an air circulation pump, a reaction
vessel, SnCl2 dispenser, an acidic gas trap and a four-way
stop-cock with tygon tubes to which is attached a ball
valve. The operations of the ball valve and the air circula-
tion pump are controlled by a microprocessor. A schematic
diagram of the system is shown in Fig. 1. During the deter-
mination, a known volume of the sample solution normally
5 ml is introduced into the reaction vessel using a micropi-
pette (15 ml). The reaction vessel is immediately stoppered
tightly and 0.5 ml of 10 % (w/v) SnCl2 2H2O in 1 M HCl
is added from a dispenser for the reduction reaction. Dur-
ing this time, air is circulated through the four-way stop-
cock to allow the mercury vapour to come to equilibrium
Fig. 1. Apparatus for mercury determination by cold vapour atomic absorption spectrophotometry (CVAAS) (Akagi & Nishimura, 1991).
855
and the acidic gases produced by the reaction also swept
into the sodium hydroxide solution. After 30 s the four-
way stopcock is rotated through 90 and the mercury
vapour is swept into the absorption cell. Response was
recorded on the strip chart recorder as a very sharp peak.
Peak heights were used for computations.
2.6. Determination of recovery
Recovery of mercury was determined by adding increas-
ing amounts of mercury to samples of two dierent sh
species which were taken through the digestion procedure.
The resulting solutions were analysed for mercury
concentration.
3. Results and discussion
The method described in this paper for the determina-
tion of mercury in sh provides a rapid, sensitive and accu-
rate system that can be used for routine analysis of sh. It
facilitates the relatively rapid (3060 min) wet oxidation of
samples (0.51 g). In addition, few reagents are required to
carry out the wet oxidation. In this digestion procedure, a
small amount of sample can be digested in a 50 ml volu-
metric ask (Pyrex) and the solution is diluted to volume
(50 ml) in the volumetric ask. This eliminates the time
consuming steps involved in the other digestion procedures
which include transfer of solution from the digestion ask
to a volumetric ask before making up to volume; and the
considerable numbers of reagents used. Such steps lead to
low recovery of mercury and or contamination. Recovery
studies were performed by spiking a sample with suitable
aliquots of 1 lg/ml standard mercury solution. Good
recoveries (94116%) of the spiked samples demonstrated
the accuracy of the method used (Table 1). In the acid
digestion/cold vapour technique, cleaning and rinsing of
856 R.B. Voegborlo, H. Akagi / Food Chemistry 100 (2007) 853858

Table 1 method has proven to be a simple, reliable, rapid method

Recovery of mercury from sh samples for the routine determination of mercury at levels as low
Sample Hg added Hg found Hg recovered % as 0.5 ng/g in sh tissue.
(ng) (ng) (ng) Recovery All the sh species analysed in this study are consumed
Selene dorsalis 0 28 by humans. Results of the total mercury in sh in lg g 1 on
(0.5 g) 0 27 wet weight basis from the coastal waters of Ghana, which
25 54 26.5 106
25 55 27.5 110
is part of the Atlantic Ocean, are presented in Table 2.
50 74 49 98 Mercury levels were determined in a total of fty-six sam-
50 74.5 47 94 ples, covering thirteen marine sh species. Mercury concen-
Pseudotolithus 0 20 tration ranged from 0.004 to 0.122 lg g 1 wet weight. All
senegalensis (0.5 g) 0 18 the samples had concentration of mercury below the
25 46 27 108 0.5 lg g 1 wet weight limit recommended by the FAO/
25 48 29 116 WHO (1972) and adopted by many countries (CIFA,
50 68 49 98
1992). Reports indicated that mercury levels in most spe-
50 70 51 102
cies of oceanic sh fall in the range of 00.5 lg g 1 wet
weight with most values close to 0.15 lg g 1 wet weight
glassware is an essential but laborious part of the analysis. (WHO, 1976). The most important exceptions to this rule
The proposed method not only reduces the amount of are swordsh, tuna sh, and halibut, whose values usually
glassware, it oers a fast and simple approach to sample range from 0.2 to 1.5 lg g 1 (FAO/WHO, 1972). Mercury
digestion and dilution. levels in skipjack, white tuna and yellown tuna caught in
The analytical response to mercury using standard solu- the Atlantic, Pacic and Indian Oceans come up to
tions prepared from HgCl2 salt was also employed to check 1.0 lg g 1 wet weight with most values ranging from 0.2
for mercury losses during the digestion. A comparison was to 0.3 lg g 1 wet weight (WHO, 1976). The results of our
made of peak heights obtained when mercury concentra- study are either in agreement or lower than the levels re-
tions of 25, 50 and 100 ng prepared from 1 lg/ml standard ported by the other authors for marine sh from other
Hg solution were taken through the digestion procedure areas of the world (Al-Majeed & Preston, 2000; Love
and the same concentrations taken directly into the volu- et al., 2003 Nixon et al., 1994; WHO, 1976).
metric asks and diluted with the diluent. There was visu- Mercury content in sh is considered to be a good indi-
ally no dierence in the two calibration curves. Ratio of the cator of human exposure to organic or methylmercury
peak heights of the digested standards to the undigested contamination. That mercury in sh appears to be predom-
standards were 9699% indicating good recoveries. Stan- inantly in the form of methylmercury has been conrmed
dard solutions prepared from mercuric chloride salt to be by many publications (Al-Majeed & Preston, 2000; Ander-
used for calibration of the analyzer could therefore either sen & Depledge, 1997; Bloom, 1992; Lasorsa & Gill, 1995;
be subjected to the digestion procedure as the samples or WHO, 1976). Therefore, diet consisting particularly of sh,
used as it is. Most digestion procedures for mercury deter- could be the main source of exposure to methylmercury in
mination employed condensers to prevent mercury losses the general population. The results of this study as such
during the heating. In this procedure, a condenser was provide a basis for assessment of human exposure to meth-
not used but excellent recoveries were obtained using an ylmercury. The concentrations of mercury in the sh sam-
open digestion technique, possibly because of the long neck ples obtained in this study are not high when compared to
of the volumetric ask allowing for reux.
Precision and accuracy of the analytical procedure were
evaluated by repeated analyses of samples and certied Table 2
1
reference material (Dogsh muscle, DORM-2) from the Mercury concentrations (lg g wet weight) in sh species samples from
the coastal waters of Ghana
National Research Council of Canada. The validity of
the method has been proved by the agreement between Species name N Mean SD
the measured (4.604.76 lg g 1) and certied (4.15 Lagocephalus lagocephalus 4 0.066 0.023
4.79 lg g 1) concentrations in the dogsh muscle Stromatteus atola 4 0.004 0.003
Brachydeuterus curitus 5 0.037 0.017
(DORM-2) Certied Reference Material. The results from Panulirus argus 5 0.035 0.015
the analysis were all within the 95% condence limit. The Calappa rubroguthata 5 0.057 0.022
use of micropipette (15 ml) for the introduction of the di- Gerres nigri 5 0.056 0.024
gests into the reaction vessel coupled with the short diges- Decapterus rhonchus 5 0.043 0.020
tion time makes it possible to analyse more than one Braehydentera aurita 3 0.122 0.030
Diplodus puntazzo 4 0.070 0.013
hundred samples a day. The judicious practice of thor- Parapristipoma humile 3 0.112 0.021
oughly rinsing all glassware with 0.5% (w/v) KMnO4 solu- Selene dorsalis 5 0.034 0.023
tion minimizes the chances for contamination from Galeoides decadactylus 5 0.041 0.020
extraneous mercury and allows the accurate determination Pseudotolithus senegalensis 3 0.031 0.025
of mercury concentrations as low as 0.01 ng/ml. This N, No. of samples; SD, standard deviation.
 R.B. Voegborlo, H. Akagi / Food Chemistry 100 (2007) 853858
some other areas of the world and can be said to reect
background mercury concentrations that are even much
lower than most published mercury concentrations in sh
from non-polluted areas of the world. For example, mer-
cury in the edible portion of various sh species landed
at Irish ports during 1993 are in the range of 0.10.39 with
a mean of 0.1 within which our values fall (Nixon et al.,
1994). These levels are reported to be low and are well
within the maximum limits set by the European Commis-
sion for mercury in sheries products. Mercury concentra-
tions reported in our study are lower by an order of
magnitude when compared to values reported for other
tropical, less industrialized areas like Indonesia, Thailand
and Papua New Guinea (CIFA, 1992). This conrms the
assertion that geographical location in addition to other
factors like metabolic dierences appears to be important
with regards to the mercury content of sh; and this is fur-
ther illustrated by the analysis of sh from dierent loca-
tions (WHO, 1976). Cod sh samples obtained from the
strait between Denmark and Sweden, which is heavily con-
taminated, had values up to 1.29 lg g 1 wet weight; cod
caught in the area of Greenland had values of 0.012 to
0.036 lg g 1 wet weight, whereas North Sea cod had values
in the range of 0.1500.195 lg g 1 wet weight. In a study of
swordsh from six areas extending from Caribbean Sea to
the Grand Banks, signicant variations from one area to
another were observed in average mercury levels. Though
the estimation of maximum amounts of daily intake of
mercury from the consumption of sh cannot be obtained
due to lack of information on nutrition survey on the pop-
ulation in Ghana, the results obtained indicate that mer-
cury content of sh from the coastal waters of Ghana is
unlikely to constitute a signicant mercury exposure to
the public because of sh consumption.
4. Conclusion
The proposed method oers a fast and simple approach
to sample digestion, dilution and mercury determination as
low as 0.5 ng/g in sh. Mercury levels determined in fty-six
samples covering thirteen species ranged from 0.004 to
0.122 lg g 1 wet weight. All the samples had concentrations
of mercury below the FAO/WHO recommended limit of
0.5 lg g 1 wet weight. These levels do not therefore consti-
tute any signicant health hazard to the general population.
Acknowledgement
The technical assistance of the sta of NIMD to one of
the authors (R.B.V.) during his visit to the institute is
highly acknowledged.
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