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Introduction 

The natural gifts of aquatic resources, fish and 
other fishery items have played an important role 
on the culture and life style of the people of 
Bangladesh. Fisheries and aquatic resources are 
economically, ecologically, culturally and aesthe-
tically important to the nation. In Bangladesh, at 
present there are 260 freshwater fish species, 12 
species of exotic fish, 475 species of marine fish 
and 60 species of prawn and shrimp available 
(Chandra, 2006). Fisheries sector contributes to 
GDP 5.2%, animal protein supply 63% and 
foreign exchange earning 4.8% for the nation 
(Chandra, 2006). Huge amount of fish catches 
from fresh and marine water during winter 

season when the natural depression attain 
shallow water and even dry up, naturally the fish 
catch attains peak.  

During this time the Bay of Bengal, the coastal 
crisscross channels and other depressions remain 
calm and quiet and as a result fishing activities 
are strengthened and huge fishes are harvested 
during this period than the other seasons. The 
fishes included shrimp, chhuri, loittya, faissya, 
popa, bailla, laukkya, puti, elgona, fewa, etc. The 
necessity to cultivate dry fish looms up when 
huge quantity of fresh fish caught everyday 
remains unsold because of shortage of customers. 
Such huge catch of fish cannot be sent to the 
towns or the metropolis on a daily basis either for 

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Screening of organochlorine insecticides (DDT and heptachlor) in 
dry fish available in Bangladesh 

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Received:                             28 July 2008 
Accepted:                            29 July 2008 
Available Online:               31 July 2008  
 
DOI: 10.3329/bjp.v3i2.997 
 
 
Cite this article: 
Bhuiyan MNH, Bhuiyan HR, Rahim 
M, Ahmed K, Haque KMF, Hassan 
MT, Bhuiyan MNI. Screening of orga-
nochlorine insecticides (DDT and 
heptachlor) in dry fish available in 
Bangladesh. Bangladesh J Pharmacol. 
2008; 3: 114-20. 

$EVWUDFW 

The concentrations of organochlorine insecticides DDT and heptachlor were 
investigated to estimate the current status of insecticides used in dry fish. The 
most popular dry fish– ribbon fish (chhuri), shrimp (chingri) and bombay 
duck (loittya) were selected for this study and these dry fishes were collected 
from different markets of Dhaka and Chittagong. The range of DDT used in 
all the samples was 3.04 to 875.0 ppb. The range of DDT in ribbon fish 131.6 to 
149.4 ppb, in shrimp 3.04 to 318.2 ppb and in bombay duck 61.9 to 875.0 ppb 
was found. The range of heptachlor used in all the samples was 0.7 to 5.5 ppb. 
The range of heptachlor in ribbon fish 1.7 to 2.3 ppb, in shrimp 0.7 to 3.8 ppb 
and in bombay duck 1.8 to 5.5 ppb was found. The heptachlor was not found 
in ribbon fish (CTG-2), shrimp (DHK-2 and CTG-1) and bombay duck (DHK-
1). The concentration of heptachlor in dry fish compare to DDT was found too 
much less.  

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shortage of transport or fish traders not willing to 
pay the right amount of money. As a result winter 
is considered to be the peak season for processing 
dry fish and continues till the onset of the rainy 
season. The dry fish is properly known as 
“SHUTKI” (in Bangla). 

For long conservation of fish by drying is 
common practice in Bangladesh. This practice is 
usually made in the remote coastal isolated 
islands and in inland depressions where chilling 
and freezing facilities are lacking. Most of the 
marine fishes which are caught in remote areas 
and islands viz., Afatiar Chor, Dublar Chor, 
Kutubdia, Khuruskul, Moheskhali, Rangabali, 
Sonadia and St. Martin’s are simply sun dried. 
The finally dried fish products are generally 
stored in a dump warehouse either at the site or 
nearby coastal towns. In addition to this, the 
weather is humid particularly during the 
monsoon period. Due to high moisture content in 
the weather and dump condition of warehouse, 
the dry fishes absorbed moisture so rapidly that 
the fish becomes suitable for infestation by beetles 
and mites. Most unexpected cause of infestation is 
that the fishermen do not dry fishes properly due 
to loss of weight i.e., the fishermen want more 
profit selling the dry fishes in weight. For 
protection of dry fish from infestation they use 
insecticides whatever they are getting within their 
reach. These insecticides are health hazard both 
for users and consumers.  

In many countries of the world like Bangladesh, a 
great deal of indiscriminate and dangerous usage 
of insecticides because of the absence of clear 
recommendations for control of insects and lacks 
of training in the nature and correct uses of 
insecticides. There are many insecticides sold in 
the markets without names and insecticides are 
not true to the label. Unknown diluted varieties 
rarely contain any instructions on heath hazards 
are sold to unaware users. Nobody can draw a 
statistic on insecticide causalities, although 
newspapers reports on deaths and sufferings or 
chronic effects are common in Bangladesh.  

DDT (dichlorodiphenyltrichloroethane) and 
heptachlor are banned in Bangladesh but there is 
no statistical figure about these organochlorine 
insecticides (UNEP, 2002). Bangladesh is party to 
the Stockholm Convention. A national 
implementation plan is already underway from 

September 2002 for the POPs (Persistent organic 
pollutant) through the assistance of UNEP. The 
Department of Environment, a member of 
Pesticide Technical Advisory Committee (PTAC) 
control the pesticide registration scheme in 
Bangladesh. No pesticides and insecticides are 
registered without reviewing the available 
Toxicological and Eco-toxicological information 
of the candidate pesticide. However, there is no 
specific legislation for controlling the production 
and use of hazardous industrial chemicals 
(UNEP, 2002). 

The most significant properties of the 
organochlorine insecticides are their extreme 
lipophilic nature and resistance to 
biodegradation, which results in their 
accumulation and concentration in fatty tissues 
and their extreme persistence in environment 
(Taqnnenbaum, 1979). Among the insecticides 
DDT is a commercial organochlorine insecticide 
that has been widely used on agricultural crops as 
well as for vector control (ATSDR, 1995). DDT 
and its by-products can persist in soil and 
sediments for more than 15 years and are known 
to bio accumulate in animal tissues. DDT had 
been banned for all uses in 49 countries and 
restricted to vector control in 23 (PANNA, 1995). 
The half-life of DDT in humans is approximately 
4 years (Noren and Meironyte, 2000). Heptachlor 
is another organochlorine cyclodiene pesticide 
that has been used to control termites and as an 
insecticide on seed grains and food crops. 
Heptachlor epoxide, the main metabolite of 
heptachlor, is extremely persistent in soil. In some 
cases, trace amounts of heptachlor epoxide have 
been found in soil 14–16 years after application 
(Extoxnet, 1996). Plants can draw heptachlor 
epoxide directly from the soil, and the chemical 
bioaccumulates in animals. It has been banned or 
restricted in more than 60 countries (WHO, 1988). 
However, some of these countries still permit its 
use for termite and other pest control, and many 
developing nations still use for agricultural 
purposes (Noronha, 1998). Despite the imposition 
of a ban on use in the United States in 1988, U.S. 
customs data showed that it was exported in large 
quantities through 1994 (Panna, 1997). A study 
showed that countries have restricted and banned 
heptachlor; levels detected in breast milk have 
dropped, often by more than 10-fold (Jensen and 
Slorach, 1991). But another studies showed that in 

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the United States, levels in the Southeast were 
nearly double the levels in the rest of the country 
during the period when it was still used (Savage, 
1981).  

The objectives of this study were to detection and 
determination of the concentration level of 
insecticides (DDT and Heptachlor) in dry fish 
used for conservation of dry fish and to elucidate 
the contamination status of organochlorine 
insecticides. By physical inspection of the sites 
(Sonadia Island, Moheskhali, Cox’s Bazaar, 
Kuakata and Dublar Chor) where the fishes are 
drying and processing by the fishermen, we 
become confirmed that some fishermen do not 
dry properly or they do not have proper 
knowledge about preservation and they used 
some poisons (insecticides) without label or label 
with improper instructions.  

 

Materials and Methods 

Sampling 

Samples from New Market (DHK-1), Karwan 
Bazaar (DHK-2), of Dhaka and Asadgonj (CTG-1), 
Reajuddin Bazaar (CTG-2), of Chittagong was 
collected. Three most popular species of dry 
fishes namely ribbon fish (Chhuri), shrimp 
(Chingri) and bombay duck (Loittya) were 
collected from each market. Total number of 
samples were 12. 

The control samples of three different fishes were 
collected from drying yards of Sonadia island that 
are known sample treated with no insecticides 
and taken into account as blank.  

Apparatus 

Mincer fish chopper (Weisser No. 81 K), Soxhlet 
extractor, separatory funnels (500 and 200 mL), 
chromatographic tube (20 mm i.d. 50 cm long), 
sample concentrator (Techne dry block DB.3), 
round bottomed flask (500 and 100 mL),  
volumetric flask (50 and 10 mL), gas 
chromatograph (GC-14B, Shimadzu), syringe (10 
µL, Hamilton Co.). 

Reagents 

Acetone, diethyl ether, dimethyl formamide 
saturated with petroleum ether, n-hexane, 
petroleum ether (30-60°C), petroleum ether (30-
60°C) saturated with dimethyl formamide, eluting 

mixture I (petroleum ether + diethyl ether 94:6 v/
v), standard solutions, eosin solution (2 mg in 100 
mL), sodium sulfate solution (2 g/100 mL NaSO4 
10 H2O),  sodium sulfate anhydrous (heated for at 
least 2 hours at 550°C), florisil 60-100 mesh 
(heated for at least 2 hours at 550°C, cool and 
stored in tightly stoppered container, prior to use 
heated for at least 5 hours at 130°C, cool and add 
5% w/w water, shake this mixture for at least 20 
min and stored in a container for at least 10 
hours), cotton wool.  

All the solvents used for the analysis purchased 
from MERCK, Germany. DDT and heptachlor 
standards were obtained from Sigma Chemicals. 

Sample preparation 

All the samples are finely comminuted in a 
mincer; heating of the samples during 
comminuting is avoided by briefly chopping 
several times (Hans and Zeumer, 1987).  

Extraction 

Triturate a sample of 25 g, with sodium sulfate to 
dry, powdery mixture, with the aid of an 
extraction thimble; extract the mixture 
exhaustively with Petroleum Ether in Soxhlet 
apparatus. Concentrate just to dryness the extract 
solution by a concentrator and dilute to 25 mL 
with petroleum ether saturated with dimethyl 
formamide (Hans and Zeumer, 1987). 

Clean up 

Clean up was done in two steps- (Hans and 
Zeumer, 1987). 

a) Dimethylformamide-petrolium ether partition: 
Transfer the solution (dissolved in 25 mL 
petroleum ether saturated with dimethyl 
formamide) to 250 mL separatory funnel. Rinse 
the flask with small portion of a previously 
measured amount of 75 mL dimethyl formamide. 
Then add the remainder of the dimethyl 
formamide to the separatory funnel, and shake 
vigorously for 1 min. Drain the dimethyl 
formamide phase, and again extract the 
petroleum ether phase with 10 mL dimethyl 
formamide. Transfer the combined dimethyl 
formamide phases to a 500 mL separatory funnel, 
and add 200 mL sodium sulfate solution. Add a 
few drops of eosin solution to achieve better 
recognition of phase separation in the subsequent 
partition. Then extract successively with a 40 mL 

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portion and three 25 mL potions of petroleum 
ether for 1 min each time. Wash the combined 
petroleum ether phases with 10 mL water, dry on 
sodium sulfate, filter through a cotton wool plug, 
add 5 mL n-hexane, and concentrate to 
approximately 5 mL. 

b) Florisil column chromatography: About half filled 
a chromatographic tube with petroleum ether, 
and sprinkle with 30 g florisil in small portions 
through a funnel with stopcock open, tapping the 
column in the process. Cover the florisil with an 
approx. 2 cm layer of sodium sulfate. Drain the 
supernatant solvent to the top of the column 
packing. Pipette the sample solution on to the 
column. Let the solution percolate to a level of 1-2 
mm above the top of the column. Then rinse the 
flask with small portions of eluting mixture I, add 
the rinsings to the column, and also let them 
percolate to a level of 1-2 mm above the top of the 
column. Next eluate the column with the 
remainder of the total 200 mL amount of eluting 
mixture I, at a flow rate of about 5 mL/min. Add 
5 mL n-hexane to the eluate, concentrate the 
eluate to 5 mL and dilute with n-hexane to 10 mL.   

Sample analyses 

The DDT and heptachlor residues were analyzed 
by GC-14B, Shimadzu with an electron capture 
detector (ECD), a manual sampler and GC 
solution software. A column of 3.1 m x 3.2 mm; 
I.D glass spiral; stationary phase silicon OV-17, 
5%, aging 300°C, support chromosorb-W-AW-
DMCS, mesh 80/100, 1 μm film thickness was 
used for the chromatographic separation of 
insecticides. The temperature was fixed for the 
injector at 250°C, column at 280°C, detector at 
280°C. The carrier gas was nitrogen with a 60 
mL/min-flow rate. 1.0 uL sample was injected for 
each run and the running time was 25 min. 
Standards’ peak were identified by injecting high 
concentration of the standard (0.5 ppm and 0.25 
ppm) and the retention time for DDT and 
heptachlor were determined. Then calibration 
was done at 3 points (25, 50 and 100 ppb) by 
composite stock standard solution. GC system 
was calibrated using external standard technique. 
Individual standard stock solution (100 mg/L) 
was prepared by weighing appropriate amounts 
of active ingredients in a brown bottle with a 
Teflon-lined screw cap and dissolving the 
weighed standard in HPLC grade hexane. Stock 

standard solution was used to prepare primary 
dilution standards. Appropriate volume of each 
individual stock solution was taken in a 
volumetric flask and mixed the solutions to 
obtain composite stock standard solution.  

Analytical quality control 

Gas chromatograph equipped with ECD was 
checked for linearity. Instrumental limit of 
detection for GC-ECD was 1.0 ug/L for 
organochlorine pesticides. An aliquot of dry fish 
samples which were collected as blank and 
treated exactly as a sample including exposure to 
all glassware, equipments, solvents and reagents 
used with the sample matrix. No analyte peak 
was detected in laboratory reagent blank. An 
aliquot of fortified samples matrix were prepared 
to which known quantities of the pesticides were 
added in the laboratory in ppb range. This 
laboratory fortified matrix was analyzed exactly 
like the sample. Extraction and clean up were 
done as mentioned and the recoveries from 
untreated control samples of dry fish fortified 
with the analyzed compounds at level of 25 ppb 
were 96-100% for heptachlor and 98-100% for 
DDT. Prior to injection of the first sample 
solution, a standard solution was injected at least 
three times to check the operating conditions and 
the constancy of the detector signals. Further 
linearity of the ECD signal was checked by 
injecting serial dilutions of DDT and heptachlor. 
A standard solution injected after at least every 
other sample solution so that any alterations of 
the gas chromatographic system recognized due 
to column contamination. 

 

Results and Discussion  

No studies of OCPs (DDT and heptachlor) in the 
dry fish of Bangladesh have been carried out. The 
present study have been undertaken in order to 
provide the preliminary information on the 
concentration of DDT and heptachlor in dry fish 
for the first time and to investigate their 
contamination level. The detected compounds 
were DDT and heptachlor. The results obtained 
are alarming for Bangladesh. All of the samples 
contained organochlorine insecticides are shown 
in Table I. The concentrations of DDT in the 
samples of ribbon fish collected from DHK-1, 
DHK-2, CTG-1 and CTG-2 were 149.4, 131.6, 140.4 

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and 137.5 ppb respectively; in the samples of 
shrimp collected from DHK-1, DHK-2, CTG-1 and 
CTG-2 were 3.04, 69.4, 3.8 and 318.2 ppb 
respectively and in the samples of bombay duck 
collected from DHK-1, DHK-2, CTG-1 and CTG-2 
were 156.6, 182.3, 875.0 and 61.9 ppb respectively. 
The range of DDT use in all the samples was 3.04 
to 875.0 ppb. Among the samples the DDT (except 
CTG-2) was found comparatively lowest in 
shrimp and highest in bombay duck. The ribbon 
fish samples of all places contained almost about 
same concentration of DDT. The level of 
concentration of DDT in dry fish is a great 
concern but more concern is such a dangerous 
poison is still using in our some popular food 
items such as dry fish though it is banned in our 
country. 

DDT is a slow poisoning substance. It can transfer 
from generation to generation through breast 
milk (Solomon and Weiss, 2001). It is classified as 
"moderately toxic" by the US National 
Toxicological Program and "moderately 
hazardous" by WHO, based on the rat oral LD50 of 
113 mg/kg (WHO, 2005). Farmers exposed to 
DDT occupationally have an increased incidence 
of non-allergic asthma (Brow, 2007). 
Organochlorine compounds in general have been 
linked to diabetes (Jones et al., 2008). EPA 
classified it as a class B2 probable human 
carcinogen (US EPA, 1987). A study of malaria 
workers who handled it occupationally found an 
elevated risk of cancers of the liver and biliary 
tract (Rogan and Chen, 2005). Exposure to it 
before puberty increases the risk of breast cancer 
later in life (Clapp et al., 2008). In areas where it is 
used for malaria control, infants can be exposed 

via breast milk in levels that exceed the W.H.O's 
acceptable daily intake value for DDT (Bouwman 
et al., 2006; Ntow et al., 2008).  

From the Table I, we found that the 
concentrations of another organochlorine, 
heptachlor in the samples of ribbon fish collected 
from DHK-1, DHK-2, and CTG-1 were 1.2, 1.9 and 
2.3 ppb respectively but in CTG-2 was not 
detected; in the samples of shrimp collected from 
DHK-1 and CTG-2 were 0.6 and 3.8 ppb 
respectively but in DHK-2 and CTG-1 were not 
detected; in the samples of bombay duck collected 
from DHK-2, CTG-1 and CTG-2 were 1.7, 5.5 and 
2.3 ppb respectively but in DHK-1 was not 
detected. The ranges of heptachlor use in all the 
samples were 0.7 to 5.5 ppb. Among the samples 
the heptachlor was found comparatively lowest in 
shrimp and highest in bombay duck. The ribbon 
fish samples of all places contained almost about 
same concentration of heptachlor. The 
concentration of heptachlor in dry fish compare to 
DDT was found too much less. In some samples 
(ribbon fish sample of CTG-2, shrimp sample of 
DHK-2, shrimp sample of CTG-1, bombay duck 
sample of DHK-1) heptachlor were not present. 
But in the chromatogram of ribbon fish sample of 
CTG-2 and shrimp sample of DHK-2 showed a 
big saturated peak which indicates a high 
concentration of organochlorine rather than DDT 
and heptachlor. That peak was not our targeted 
peak. But it can indicate that there is another 
organochlorine which is used in high 
concentration like DDT. It is also alarming to 
Bangladesh that a banned item like heptachlor is 
still using in dry fish as well as DDT and it can 
assume from the study that a composite mixture 

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Table I 

Summary of DDT and heptachlor concentrations in the dry fish samples   

Dry fish   Insecticides   

DHK-1               DHK-2           CTG-1              CTG-2  

Ribbon fish Chhuri) DDT 149.4 131.6 140.4 137.5 
Ribbon fish Chhuri) Heptachlor 1.7 1.9 2.3 ND 

Shrimp (Chingri)  DDT 3.04 69.4 3.8 318.2 

Shrimp (Chingri) Heptachlor 0.7 ND ND 3.8 

Bombay duck (Loittya) DDT 156.6 182.3 875.0 61.9 

Bombay duck (Loittya) Heptachlor ND 1.8 5.5 2.3 

The concentrations are in ppb unit; ND = Not detected  

                        Name of locations  



 

 
of insecticides is used for the conservation of dry 
fish. Heptachlor has been shown to 
bioaccumulate in fish and cattle. Most of the 
heptachlor that is swallowed is absorbed into 
blood. Heptachlor can pass directly from a 
mother's blood to an unborn baby through the 
placenta (LDWG, 2007). Animals fed heptachlor 
throughout their lifetime had more liver tumors 
than animals that ate food without heptachlor. 
EPA has classified heptachlor as a probable 
human carcinogen (B2) and established an oral 
cancer slope factor of 4.5 per mg/kg-day (LDWG, 
2007). 

Almost all industrialized countries and many 
developing countries around the world have 
recognized the hazards of persistent pesticides 
and have banned them. But in some developing 
countries, POPs insecticides are readily available 
in spite of official bans or severe restrictions. In 
those countries a government may lack the 
resources and infrastructure to implement and 
enforce the legislation fully. People selling these 
chemicals in isolated villages may have no idea 
that sale or use of the pesticides is restricted.  

The government of Bangladesh should take all the 
necessary steps to combat the situation, it can be 
the steps to implement the legislation and 
improving the awareness of the related people 
through some program, public education 
campaigns and announcement of harmful rule of 
those insecticides and make difficult the 
availability of those insecticides in market either 
its true pack or false pack. The stocker should dry 
correctly and should pack carefully so that the 
fish can not absorb moisture in monsoon. The 
other way of preservation can follow such as 
freezing, canning and curing. From the study, we 
can say that, the stocker used insecticides 
abundantly. They use DDT as a compulsory 
preserver of dry fish. Other insecticides are also 
used as a composite mixture of insecticides to get 
good preservation.  

Due to our limitation of facilities, we could not 
study about other organochlorine and 
organophosphorus but we suspect that there are 
other organochlorine insecticides contaminations 
in the dry fish, as our chromatograms showed 
some unexpected peaks in every sample. 

Acknowledgements       

This study was done under the research and 
development project of Biochemistry Division, BCSIR 
Laboratories, Chittagong. The authors are indebted to 
Mr. K.K. Nath, CSO, BCSIR, Chittagong; Dr. Mamtaz 
Dawlatana, CSO, IFST, BCSIR; Mr. Jasim Uddin 
Chowdhury, CSO, IFRD and Mrs. Sayeda Shahida 
Begum, PSO, IFST, BCSIR. The authors also wish to 
thank Mrs. Aysha Akter, Lecturer, Institute of Marine 
Sciences, Chittagong University for helping in 
identification of fish.  

             

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Md. Nurul Huda Bhuiyan (Principal contact) 
e-mail: nhbmb@yahoo.com  

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