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Journal of National Oceanographic, Atmospheric and Maritime Institute

©NOAMI, Eastern Plaza, Dhaka, Bangladesh
Vol.22, No.2, 1- 8, 2005
ISSN: 1027-2119

DETERMINATION OF ARSENIC, CHROMIUM, SELENIUM AND ZINC IN
SOME TROPICAL FISH BY NEUTRON ACTIVATION

A.K.M.Sharif
Bangladesh Atomic Energy Commission, Dhaka, Bangladesh

K.R.Krishnamoorti
Analytical Chemistry Division, Bhava Atomic Research Center, Trombay, India

M.Alamgir and Stanley A.Bajue
Department of Physical, Environmental and Computer Sciences
Medgar Evers College, CUNY, NY, USA
____________________________________________________________________________________________________________

ABSTRACT

Determination of arsenic, chromium, selenium and zinc in several commonly consumed tropical
marine fishes have been by neutron activation followed by radiochemical separation to remove
the interfering activities of sodium, potassium, bromine, and phosphates, etc., in order to
establish the baseline data and to measure the levels of contamination, if any. The results of this
study positively indicate that the marine fishes of Bangladesh have concentrations much below
permissible levels for these toxic levels for these toxic elements. A radiochemical scheme for the
separation of these trace elements in biological materials is also presented in this paper.

Key words: Arsenic, Chromium, Selenium, Zinc, Tropical Fish, Tropical Fish, Bay of Bengal,
Bangladesh
_______________________________________________________________________

INTRODUCTION

Determination of essential and toxic trace elements in biological materials, especially foodstuffs,
has been become the need at this present time. Earlier workers [1-3] have reported that trace
metal contents of certain foods, especially fish can be contaminated with industrial effluents and
wastes discharged into oceans, lakes, coastal waters, etc. Various species of fish [4-6] may
uptake heavy metals in their tissue, which may in some approach toxic levels.
J.Nat.O.A.M.Institute, Vol.22, No.2,1-8,2005

Some of the investigators [7-12] have reported significantly high levels of cadmium and arsenic
in some species of fish. However, the investigation of DE GOEIJ [13] and GUINN [14] have
shown that there is no significant difference of these levels of certain trace elements, like arsenic,
cadmium than those in unpolluted areas.

Whether or not fish will be contaminated will depend on the chemical form of the element and
its concentration in the surrounding medium, microbiological activity in the marine environment,
texture of the sediment, type and age of the fish, etc. However, there are still insufficient data
available in Bangladesh for toxic metals like arsenic, cadmium, etc. It is strongly believed that
this study will consequently be a great help for Bangladesh’s economy in view of quality
assurance for trade as well as the health, safety and benefit of her people. This paper presents
information on the concentration of arsenic, chromium, selenium and zinc in some varieties of
fish and also describes briefly the chemical procedure followed.

The instrumental neutron activation analysis (INAA) of such elements as As, Cr, Cd, Zn and Cu
in animal organs and fish tissues is difficult because of the 24Na and 86Br activities developed on
irradiation. This is because the photopeaks of radioactive products of these elements are masked,
particularly by the Compton continuum of the high 24Na matrix activity, thus posing problem not
only in handling but also in the computation of the peak areas from the γ-ray spectra. This
problem has necessitated post-irradiation chemical separation of isotopes of interest. This paper
describes such a scheme for determination of seven trace elements (As, Cr, Se and Zn) in
biological materials.

EXPERIMENTAL

Samples collection and irradiation. Eight varieties of common marine fish, selected in
accordance with their public flavor for Bangladeshi (near coastal belt) in both taste and cost,
namely, Coilia neglecta, Cirrhinna reba,Johnius argantus, Harpondon nehereus, Setipinna
phasa and Lepturacanthus savala were collected from the coastal belt of the Bay of Bengal and
sun-dried after removal of their internal organs, head, skin, and tails. The dried samples were
then chopped into pieces with the aid of a stainless steel knife (steam cleaned). Only the edible
muscle tissue samples were used for analysis. The sample pieces were dried at 105o-110oC in an
oven until a constant weight was obtained (dry weight).

The dried samples were ground, sieved and thoroughly mixed in a stainless steel rotating drum
for 100 hours to produce a homogeneous powder. The sample powder was finally preserved in
clean and dry polyethylene bottles. Portions of the samples (200-300 mg each) were heat-sealed
in polyethylene bags and irradiated along with a known amount of MA-A-2 ™, the fish flesh
homogenate standard of IAEA in the CIRUS reactor at Bhabha Atomic Research Center,
Trombay, Bombay, India, at a flux of about
(0.5 to 1) .1012 n. cm-2. s-1 for 20 hours.
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Chemical reagents. (1) NH3, HCl, HNO3, HClO4 and acetic acid; (2) Na2SO3 solution:
4 mg/cm3 ; (3) NaOH pellets; (4) thioacetamide; (5) H2O2; (6) BaCl2 (=0.2M solution in water;
(7) NH4H2PO4= 1M solution; (8) hydroxylamine hydrochloride (NH2OH.HCl).

Dissolution. Each of the irradiated samples was allowed to “cool” for 4-5 days to enable the
decay of short-lived isotopes and also to reduce the 82Br and 24Na activities. About 10 mg carrier
for each of the element was added to a 100 ml round-bottomed flask and the irradiated sample
was carefully opened and emptied into it. Then, the sample was digested in Bethge’s apparatus
with a mixture of 6 cm3 concentrated nitric acid and 2 cm3 of concentrated perchloric acid till a
liquid remained in the flask.

Chemical separation and determination of metal

After the digestion was completed, the distillates were, evaporated to dryness on a sand bath to
remove nitric acid, and leached with conc. HCl (10 cm3) and preserved to be combined with the
filtrates from chromium and selenium precipitation.

(1) Determination of chromium: 4 cm3 HClO4 was added to the residue in the flask and drops
of conc. HCl added to the hot solution to distil of CrO2Cl2 . The distillate was collected in
10 ml sodium hydroxide solution (1N) . To this was 1.5 cm3 BaCl2 solution followed by
addition of 2-3 drops of H2O2. The pH was set at 6. BaCrO4 was digested in a water bath
and was filtered, dried and counted for 51Cr. The filtrate was combined with the distillate
from decomposition step.
Cr is oxidized to Cr(VI) by HClO4
H2Cr2O7 + 4 HCl → 2CrO2Cl2 + 2H2O(reddish vapor)
CrO2Cl2 + 2NaOH →Na2CrO4 +2HCl (yellow distillate)
Na2CrO4 + BaCl2 → BaCrO4 + 2 NaCl
(2) Determination of selenium: To the residue remaining in the flask 20 ml of conc. HCl was
added and the volume kept at 50 ml. To the warm solution 4-5 cm3 Na2SO3 solution was
added and digested in a water bath for 1 hour. The precipitated Se metal was filtered,
dried and counted for 75Se. This filtrate also was combined with the distillate from the
decomposition step.
MCl2 + SO2 +2H2O → M (metal) + 2HCl + H2SO4
(3) Determination of arsenic: The acidity of the first distillate, with the filtrates from
chromium precipitation and selenium precipitation mixed together, was adjusted to 1M
HCl and 1 ml of a 1% solution of thioacetamide was added to the boiling solution. This
was digested in a water bath. Precipitates of sulfides of As were filtered off, washed,
dried and counted 76As.
CH3CSNH2 + H2O → CH3CONH4 + H2S
CH3CONH2 + H2O→ CH3COONH4
H2S + M+++ → MS + 2H+
[M = As]
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(4) Determination of zinc. The filtrate from (3) was evaporated to dryness in a boiling water
bath followed by addition of drops of conc.HNO3 until the ammonium salts were
completely destroyed. To this was added 1 cm3 1M NH4H4PO4 solution. The pH of the
solution was then adjusted to 6 and it was heated for a few minutes. The mixture was then
kept in a water bath for 1 hour. The precipitates obtained were filtered off, washed, dried
and counted for 65Zn.
ZnCl2 + NH4H2PO4 → ZnNH4PO4 + 2HCl [M= Zn]

Counting
The samples and the standards were counted on a 45 cm3 HPGe detector connected to a 4096
channel pulse-height analyzer (Ortec PCA-MCA card). The energies (in keV) chosen for the
evaluation of the peak areas were: 76As(26.4 h), Eγ=657 keV (since, the photopeaks of 122Sb
(561keV) interfered with photopeak 76As at 559 keV, the 657 keV photopeak was chosen to
measure 76As photopeak area) ; 51Cr(27.7 d), Eγ=320.08keV; 75Se (119.77 d), Eγ = 264.66
keV; 65Zn (243.9 d), Eγ= 1115.5 keV.

Accuracy and precision
Experiments were initially carried out using radioactive tracers and the corresponding
carriers to evaluate the recoveries. The yields were in the range of 93% to 98%. The accuracy
of the method was evaluated by analyzing homogenate fish flesh (IAEA) Standard Reference
Material, MA-A-2™. Our results (values in µg/g) are in good agreement with the IAEA
certified values.

RESULTS AND DISCUSSION

The As, Se, Cr and Zn concentrations determined in the fish species are presented in
Table 1. The values are expressed in µg.g-1 dry weight. The range of concentrations found in
the fish samples are As (2.843-3.920), Se (2.961-6.274), Cr (0.498-1.8430), and Zn (37.59-
101.19). These variations are likely to be due to the migratory nature and feeding habits of
the different species of fish.

Normally, the levels of As (3 ng/ml), Se (0.09 ng/ml) and Zn (0.01 ng/ml) have been found
to be quite low in sea-water [15], but the levels of these metals as found in fish samples
under investigation are higher. This is due to the tendency of various species of fish to
concentrate certain elements in the tissue more than surrounding medium. The mean
concentrations of arsenic, selenium, chromium and zinc in fish are 3.234 µg/g, 4.385 µg/g,
1.007 µg/g and 59 µg/g, respectively.

Using neutron activation, HAMILTON AND MINSKI [16] reported the mean values for As
in fish as 2.0±0.08 µg fresh weight. Up to 174 ppm has been found in prawns from the
coastal waters of Britain [17] and 42 ppm in shrimp from the southeastern
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coastal waters of the United States [18]. Fish flour for human consumption has been
reported [19] to contain 1.8 ppm Se and tuna meal median levels as high as 5.1 and 6.2 ppm
(dry basis)[20]. High natural Se levels in tuna [21] and marine mammals [22]
have been reported. Open ocean water contains as little as 10 µg/l of Zn at the surface [23]
although coastal seawater usually contains 0.5-2 µg/l of Zn as a result of river inputs and
sewage outfalls [24-26].

Table 1
Arsenic, selenium, chromium and zinc in different varieties of fish
(µg. g-1 dry weight basis)
Types of Weight Organic
fish Analyzed, Arsenic Selenium Chromium Zinc Matter,
g %
Colia 0.36220 3.025 4.751 1.843 80.48 79.75
neglectic
Cirrhina 0.31093 3.076 2.961 1.287 49.55 84.85
reba
Johnius 0.32212 3.920 3.856 0.550 37.59 83.45
argantus
Harpodon 0.27691 3.070 6.274 0.871 101.19 81.82
nehereus
Setipinna 0.31377 3.469 3.839 0.995 50.73 82.68
phasa
Leturacan- 0.36220 2.843 4.627 0.498 38.02 81.73
Thus
salala
Stromateus - 2.52 - - 51.0 -
sinensis
Rita rita - 3.78 3.15 - - -
Mean - 3.234 4.385 1.007 59.59 82.36
Range - 2.843- 2.961- 0.498- 37.59- 79.75-
3.920 6.274 1.843 101.19 84.85

Taking 6 g of fish as the maximum consumption [27] per person per day for Chittagong and
coastal areas of Bangladesh, it is estimated that the average intake of arsenic, selenium,
chromium and zinc through fish is 3.881 µg, 5.262 µg 1.208 µg and 71.51 µg, respectively.
The daily intake of arsenic is quite low[28]. Table 2. gives the comparison of the levels of
arsenic, selenium, chromium and zinc in fish by various researchers in different countries.
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CONCLUSIONS
The present study indicates that the radiochemical separation scheme should be used
for the isolation nuclides with mutually interfering γ-rays (e.g., 75Se, with 203Hg at 265 keV )
. In the case of 76As, due to the interference of 122Sb, at 559 keV, we have chosen the less
sensitive 657 keV peak for computation of arsenic.

Eight tropical marine fish species collected from the Bay of Bengal were analyzed in order to
assess the level of trace toxic elements in this food item consumed by the
Table 2
Trace element concentrations (mg. Kg-1 wet weight ) in muscle tissue of various species of
fish from different areas of the world
Source Area Cr As Se Zn Species
Bebbington Australia, - 0.2-2.2 - 4.24- Commercial
Et al (1977) N.S.W 6.60 Species
[29] including
flathead,mullet
Roth and Israel 2.8-4.9 - - 14.9-25.5
Harnung
(1977) [30]
J.H.Powell Papua - 0.4-3.5 - 3.0-4.5 8 species
et al. New including
[31] Guinea sharks,travelly
Khan et.al Dhaka, - 2.52-5.53 3.15 26.0-93.8 6 species
(1987) Bangladesh including prawn,
[32] rita,gar
fish
Anand.S.J.S. Bombay, - 0.069- - - 6 species
(1978) India 0.931 (pampus
[33] argentius,
harpodon
neherer etc.)
*Present Bay of 0.50- 2.84-3.92 2.96- 37.59- 6 species (coilia
study (on Bengal, 1.84 6.27 101.19 neglecta,cirrhina
dry weight Bangladesh reba, johnius
basis) (1998) argentus,
harpodon
nehereus,setipina
fhasa and
lepturacanthus
savala)

*The present data were calculated on dry weight basis
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population of Bangladesh. The results indicate that the concentrations of these elements are
much below the toxic levels.

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