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Naphthalene, a Polycyclic Aromatic Hydrocarbon, in the Fish Samples from the Bangsai River of Bangladesh by Gas Chromatograph–mass Spectrometry

M. Amzad Hossain
Biotechnology Research Institute, Universiti Malaysia Sabah, Locked Bag No. 2073, 88999 Kotakinabalu, Sabah, Malaysia

Farida Yeasmin
Department of Chemistry, University of Dhaka, Dhaka 1000, Bangladesh

S.M. Mizanur Rahman
Department of Chemistry, University of Dhaka, Dhaka 1000, Bangladesh

S. Rana
Chemistry Division, Atomic Energy Centre, GPO Box 164, Ramna, Dhaka 1000, Bangladesh

Abstract/ Executive Summary:

Naphthalene, a polycyclic aromatic hydrocarbon (PAH), was detected and quantified in the selected varieties of fishes collected from the Bangsai river, one of the contaminated rivers located at Savar near the Dhaka Export Processing Zone (DEPZ), Bangladesh, during the period October 2009. Naphthalene, a carcinogenic compound, was analyzed by GC–MS as it was in the mixture of dichloromethane–hexane (1:1) crude extract of the flesh of fish samples collected from the aforesaid river. A suitable and reliable procedure for the extraction of naphthalene from the fish sample has been developed. A multi-layer clean-up (silica gel) column was used, followed by glass fiber filter (GFF) paper to eliminate the interfering organic compounds as well as the lipids and fat. It was observed that PAHs deposition on the samples takes place in different morphological parts of the biological materials. The PAH, naphthalene, was found in almost all of the fish samples and the concentration of which was in the range 0.030–1.004lg/g. Recovery studies with fortified samples indicated that the recovery efficiency for naphthalene was about 79.14%. This concentration is with in the range of values reported for other comparable regions of the world.

Keywords: Quantification; Carcinogenic naphthalene; Fish samples; GC–MS; Bangsai river; Bangladesh

Location: Bangsai river, Savar, Dhaka, Bangladesh

Start Date: 00-10-2009

End Date:

Program Area: Chemical Analysis

Commodity: Fish

Objectives:

The objective of this work is to check the toxic polycyclic aromatic hydrocarbons in the crude extract isolated from the fish sample by GC–MS.

Methodology:

Chemicals dichloromethane, hexane and other solvents used in this experiment were of HPLC grade. Anhydrous sodium sulfate (Merck,Germany) was cleaned by heating at 200°C before use. Silicagel (60–120 mesh, Merck, Germany) was activated at 400°C for 12 h prior to use. Glass fiber filter paper (Merck, Germany)was used for removal of fats and lipids. Naphthalene (Sigma–Aldrich) was used as the standard in the present study. 2.2. Fish samples: Seven different varieties of fish samples were collected from the Bangsai river, Savar, Dhaka, Bangladesh, in October 2009 and initially identified by morphological features and data base present in the library at the herbarium of the Department of Biology, University of Dhaka, Dhaka, Bangladesh. 2.3. Isolation and preparation of crude extracts: Fish samples were at first washed with tap water and then with de-ionized water. The washed samples were flayed to collect flesh, which again was washed by de-ionized water to remove blood, dusts and any other foreign particles. The collected flesh samples were ground by mortar pestle. The paste samples (5 g) were extracted three times with dichloromethane–hexane (1:1)(40 ml·3) at 80°C for 30 min. It was then filtered by glass fiber filter paper and the filtrate was evaporated near to dryness by Kuderna-Danish evaporator. 2.4. Clean-up procedure: The clean-up column (i.d. = 1 cm) was filled with cotton in the bottom. An activated silica gel (17 g) soaked with dichloro-methane was loaded into the clean-up column (5 cm), which was then topped with 1.5 cm of anhydrous sodium sulfate. Five milliliters of dichloro-methane was added to wash the sodium sulfate and the silica gel. The dried 1 ml sample was then transferred into the column; the vessel was rinsed twice with 2 ml dichloro-methane, which was also added to the column. Sixty millimeters of acetone was added to the column and allowed to flow through the column at a rate of 3–5 ml/min, and the effluent was collected. The collected effluent from the clean-up procedure was reconcentrated to 0.5 ml with K-D concentrator. 2.5. GC–MS analysis and program: The GC–MS analysis of the crude extract of fish samples was performed using a Varian GC–MS (Model Varian CP 3800) equipped with a VF-5 fused silica capillary column (30 m·0.25 i.d., film thickness 0.25lm). For GC–MS detection, anelectron ionization system with ionization energy of 70 eV was used. Helium gas was used as a carrier gas at a constant flow rate of 1 ml/min. Injector and mass transfer line temperatures were set at 250 and 280°C, respectively. The oven temperature was programmed from 50 to 200 at 8°C/min, and then held isothermal for 20 min and finally raised to 300°Cat10°C/min. Diluted samples (1/100, v/v, in methanol) of 0.2ll were manually injected in the split less mode. Identification of compounds of the crude fish extract was based on GC retention time on VF-5 capillary column, computer matching of mass spectra with standards (Mainlab, Replib and Tutorial data of GC–MS systems) and, whenever possible, by coinjection with authentic compounds (Lawless, 1995).The system suitability of the method was evaluated by the intra- and inter-day precision and accuracy of replicates. The accuracy was evaluated through recovery studies by adding known amounts of the standard solution to the extract. The recovery experiment was performed at three different standard concentrations. 2.6. Preparation of standard: Calibration graphs for the samples treated according to the described analytical procedure were made using the SIM mode.Different concentrations of naphthalene (0.625, 1.25, 2.5 and 5lg/ml) were used for calibration curve.

Source of Information: Arabian Journal of Chemistry (2014)7, 976–98

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

From the experiment, it is concluded that six fish samples such as baim, bata, puti, chapilla, prawn and taki were contaminated by various aspects such as industrial effluents, air, high-way vehicle exhaust, and highway tar samples. The findings thus suggest that awareness should be grown among the Bangladeshi nationals who live nearby the Bangsai river for not to eat these naphthalene biased fishes. The experiment also reveals the fact that the most probable source of PAHs is oil contamination originating from the effluents from different industries such as tannery, dye, plastic, chemical, and fertilizer or spillages and/or heavy ship traffic. However, the high concentration of carcinogenic naphthalene encountered in these fishes should be considered seriously as it is hazardous to human health.

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