Determination and Comparison of the Organochlorine Pesticide Residue Levels Among Benni Fish of Shadegan, Mahshahr and Susangerd Cities, Khozestan Province in Iran
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Jundishapur Journal of Natural Pharmaceutical Products 2011; 6(1): 24-31 Jundishapur Journal of Natural Pharmaceutical Products DETERMINATION AND COMPARISON OF THE ORGANOCHLORINE PESTICIDE RESIDUE LEVELS AMONG BENNI FISH OF SHADEGAN, MAHSHAHR AND SUSANGERD CITIES, KHOZESTAN PROVINCE IN IRAN Arzi A1,2, Hemmati AA1,2, Nazari Khorasgani Z2* 1Physiology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 2Department of Pharmacology & Toxicology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Received: January 2011 Accepted: June 2011 Abstract Concentrations of hexachlorocyclohexane isomers (β,γ,δ HCH), dichlorodiphenyl trichloro ethane (pp, opDDT) and its metabolites ( ppDDE, ppDDD), aldrin, dieldrin, heptachlor , heptachlor epoxide, endosulfan isomers (α, β) and metoxychlor were determined in Benni fish collected from Shadegan, Mahshahr and susangerd cities at Khozestan province in Iran. All the collected fish were contaminated by 14 investigated organochlorine pesticides. The highest and lowest mean concentrations of organochlorine pesticides belonged to β- HCH (65.36 μg/kg) and op DDT (0.13 μg/kg) and were found in Mahshahr and Shadegan Benni fish respectively. Keywords: Organochlorine pesticide, Contamination, Benni fish. Introduction Intensive use of organochlorine pesticides also been demonstrated in humans. for agriculture and health purposes in Measurable levels of organochlorine developing countries, has led to pesticides have been found in human widespread pollution of the environment adipose, tissues, blood and breast milk (1-6) Owing to their properties, including throughout the world(12-15). Nowadays low volatility, chemical stability, an extensive attention is paid by FAO environmental resistance, lipophility and (Food and Agricultural Organization and slow metabolic degradation, WHO (World Health Organization) on bioaccumulation and bioconcentration has pesticide residues in food, environment , occurred in birds and mammals and the and the possibility of harm to human food chain (2, 7, 8). following long – term of low levels Investigation a variety of wildlife has also exposure and consequent effects of clearly demonstrated that extensive chronic poisoning. Valuable information contamination of fish, birds and have been reported for organochlorine mammalian species by these compounds pesticide residue levels in environment has emerged (1, 9-11). Contamination has and food in developing countries, but *Email: [email protected] Arzi A/ JJNPP 2011; 6(1): 24-31 sufficient data about these pesticides in east by Chahar Mahal and Kohgilooyeh, Iranian fish, are very rare. Benni fish in the west by Iraq and the south by (Barbus Sharpeyi) belonged to Cyprinidae Persian Gulf. (Fig. 1) The largest rivers in family, Actinopterygii(ray finned fishes) this province are Karoon, Karkheh, Dez, class. This fish live in western Asia, Maroon and Arvand. The reedy marshes endemic to Tigris Euphrate basin. Due to called Hoors are numerous, the most importance of the problem for Iranian important of which are Hoor OL-azim and Health Officials, the organochlorine those located in Susangerd and Shadegan pesticide residue levels [(β, γ, δ- HCH), regions. Khozestan consists of 15 cities. pp, op-DDT and its metabolites (pp'DDE, Ahvaz is the provincial capital and pp'DDD), aldrin, dieldrin, heptachlor, following are the other cities: Abadan, heptachlor epoxide, endosulfan isomers Andimesshk, Izeh, Baghmalek, Bandar-e (α, β) and metoxychlor] were determined Mahshahr, Behbahan, Dezfool, Dasht-e- in Benni fish of Shadegan, Mahshahr and Azadegan, khorramshahr, Masjed-e- Susangerd cities at Khozestan province in soleyman, Ramhormoz, Shadegan, Shush, Iran. Omidieh, Aghajari and Shooshtar. The dams of Dez, Karoon and Karkheh Materials and Methods provide water for irrigation, Animal Areas under study husbandry-particularly sheep, goat and Khozestan situated in the south-west of buffalo and fishing are of high Iran, this province covers an area of 64236 importance. km2 It is bounded on north by Lorestan the Fig. 1: The map of Khozestan province 25 Determination and comparison of the Collection of fish From each of three areas under study monthly, for six consecutive months. Fish (Shadegan, Mahshahr and Susangerd ), were gray color and 300 -700g weight. 20 Benni fish were taken with fishing net (Fig. 2) Fig. 2: Benni fish Laboratory procedures All glassware used in the analysis were ml saturated NaCl solution and 100 ml carefully washed with water- detergent petroleum ether .CH3CN phases from each and rinsed with distilled water, ethanol, of 3 additional partitioning was passed acetone and pesticide grade n- hexane through same 15 ml petroleum ether in respectively. The reagents and chemicals 125 ml separator , each time shaken used were highly pure and obtained from vigorously and combined CH3CN extracts various branches of Merck Company. In in the 1 L separator. Then aqueous layer order to investigate the status of drained into another separator, and 100 ml qualification and quantification of petroleum was added to that and shaken contamination of Benni fish of Susangerd, vigorously. When layers separated, Mahshahr and Shadegan by selected aqueous layer discarded and petroleum organochlorine pesticides[(β,γ&δ- )HCH , ether layer was combined with that in pp', op'-DDT and its metabolites (pp'DDE original separator. Then, petroleum ether ,pp'DDD), aldrin, dieldrin, heptachlor , layer was washed with two 100 ml heptachlor epoxide, (α, β) endosulfan portions water. Petroleum layer was drawn isomers and metoxychlor)], different parts off through sodium sulfate and of each 20 collected fish from each of area concentrated about 10 ml by rotary was taken, grounded by and thoroughly evaporation for Florisil column mixed. 20 g of grounded fish was dried chromatography cleanup. Florisil column with anhydrous sodium sulfate and soxhlet chromatography cleanup was used for extracted with n- hexane for 8h. The fractionation of organochlorine pesticides extract was concentrated to dryness by from fish samples by elution with solvents rotary evaporation. The lipid content of of increasing polarity. The sample was dissolved in 15 ml petroleum chromatographic columns (50 cm ×1.3 ether and transferred into 125 ml I.D) were slurry packed with 10 g of separatory funnel. Then 30 ml CH3CN Florisil (activated at 675 ˚C). saturated with petroleum ether was added Approximately 0.5 cm anhydrous sodium to that separator, shaken vigorously 1 min, sulfate was placed at the top of the when layers separated, CH3CN phase was column. The column was pre eluted with drained into second 125 ml separator sufficient amount of petroleum ether. A containing 15 ml petroleum ether, shaken 250 ml round bottomed flask was placed vigorously 1 min, after layers became under the column. Then extracted separated, CH3CN phase was drained into petroleum ether was transferred into the 1 L separator containing 650 ml H2O, 40 column and allowed it pass through at 5 26 Arzi A/ JJNPP 2011; 6(1): 24-31 ml/ min , and eluted with 200 ml in the winter of 1943-1944(16), the petroleum ether- diethyl ether (94: 6) control of river blindness (onchocerciasis) (fraction 1) .The receiver was changed and in the West Africa by killing the insect eluted with 200 ml petroleum ether - vector with temephos (17) and the control diethyl ether (85: 15) (fraction 2). Each of of malaria in Africa, the Middle East and elution was concentrated to dryness by Asia with a variety insecticides are all rotary evaporation and residue was medical miracles accomplished by dissolved in 10 ml n-hexane for gas pesticides that have been documented. chromatography analysis. The separation Despite the current dispute over pesticide and quantification of pesticide residues use and the presence of low levels of their were performed on a Varian 6000 gas residues in food, groundwater, biota and chromatograph equipped with an electron air, these agents comprise integral capture detector (63 Ni) using a capillary components of our crop and health column of 25 m length and 0.53 mm protection programs. As long as they diameter contained 3% OV17 on a 60-80 continue to be used, accidental and/ or mesh chromosorb. The operating intentional poisoning of wildlife domestic temperatures were: injector 230 ˚C, stock, human and contamination of detector 320 ˚C and oven temperature was environment and food can be anticipated. maintained at 170 ˚C for 8 min, then Due to importance of the problem for programmed at 5 ˚C /min to 200 ˚C and Iranian Health Officials, FAO and WHO held at 200 ˚C for 14 min. Nitrogen gas the residue levels of organochlorine was used as carrier gas at flow rate 1 ml/ pesticides were studied in this study. min. The sample size was 1 μl. Recoveries Tables 1, 2 and figure 3 shows the results ranged from 94 to 99% and residues levels of this research. From viewpoint of were not corrected for recovery values. organochlorine pesticide concentrations, Residues of pesticides in fish were this survey revealed this fact that all of identified and quantified by comparing fish samples, in some way were their retention times and area under the contaminated by investigated pesticides curves obtained for the chromatograms of (tables 1 and 2). According to results of samples with those obtained for the this study, table1 shows the range of chromatograms of selected pesticides concentration values of each investigated standards which were analyzed under organochlorine pesticides. The