Investigation of pesticide residues in water,sediments and fish samples from Tapi River,India as a case study and its forensic significance

Abstract

This research is a case study on detection of pesticides in river water, sediment as well as fish samples from Tapi River, among the major rivers of Gujarat, India. To investigate the misuse, concentration level and occurrence patterns of persistent pesticides, samples were collected from the river. Chlorpyrifos, methyl parathion, hexachlorocyclohexane (HCH), dichloro diphenyl trichloroethane (DDT) and endosulfan were analyzed by gas chromatography technique with flame ionization detector (FID). Scanty reports are available, but after 1999, no such data are reported as some of these pesticides have been banned. Although these pesticides are still in use which we observed from the obtained results. In this river, the amount of endosulfan, chlorpyrifos, and methyl parathion was observed in surface water with concentrations of 37.56?µg/L, 0.86?µg/L and 0.43?µg/L, respectively. Endosulfan, DDT and methyl parathion detected in sediment were 38.38?ng/g, 0.65?ng/g and 0.77?ng/g, respectively. In fish samples, levels of endosulfan, chlorpyrifos, and methyl parathion detected were 101.28, 0.392, and 3.49?ng/g correspondingly. Results showed that highly toxic pesticides are still being used in the surrounding area, and there is an urgent need for enforcement of rules to control the production and application of such pesticides.     

Effects of atrazine on acetylcholinesterase activity in midges (Chironomus tentans) exposed to organophosphorus insecticides

Acetylcholinesterase activity was determined for midge larvae (Chironomus tentans) exposed to either organophosphorus insecticides (OPs) alone or OP insecticides in binary combination with atrazine (200 μg/l). Although atrazine by itself did not reduce the level of acetylcholinesterase activity, atrazine in combination with chlorpyrifos significantly decreased acetylcholinesterase activity as compared to chlorpyrifos only treatments. Although similar trends existed for malathion and methyl parathion, differences were not statistically significant. These results match previously published toxicity data where atrazine, although not acutely toxic even at much higher levels, decreased EC₅₀ values for chlorpyrifos by a magnitude of 4, decreased methyl parathion values by a magnitude of 2, and did not decrease values for malathion.     

Biodegradation of chlorpyrifos by either single or combined cultures of some soilborne plant pathogenic fungi

Abstract

Biodegradation of chlorpyrifos was studied in liquid culture media amended with either single or combined eight different plant pathogenic fungi isolated from the continuous cropping wheat fields. The average recovery of chlorpyrifos from the liquid media was found to be 86.1%. The detection limit of chlorpyrifos by the analytical method used was 19 ppb. Data showed that the growth of mixed fungi at concentrations up to 200 ppm of chlorpyrifos was higher than in the control treatment. Chlorpyrifos concentrations declined in the medium of combined fungi more than it did in the medium of any single fungus with increase in the incubation period. The amount of chlorpyrifos recovered was 79.8 ppm (39.9%) in the combined fungal cultures after 21 days. However, those recovered from the media of Fusarium graminearum, F. oxysporum, Rhizoctonia solani, Cladosporhim cladosporiodes, Cephalosporium sp., Trichoderma viridi, Alternaria alternata, and Cladorrhinum brunnescens, ranged from 48.0 to 74.8%. The half‐life value (T_1/2) for chlorpynfos was 15.8 day in the medium amended with mixed fungi. However, for the single cultures it ranged from 19.3 to 33.0 day.     

Effects of atrazine on acetylcholinesterase activity in midges (Chironomus tentans) exposed to organophosphorus insecticides

Acetylcholinesterase activity was determined for midge larvae (Chironomus tentans) exposed to either organophosphorus insecticides (OPs) alone or OP insecticides in binary combination with atrazine (200 μg/l). Although atrazine by itself did not reduce the level of acetylcholinesterase activity, atrazine in combination with chlorpyrifos significantly decreased acetylcholinesterase activity as compared to chlorpyrifos only treatments. Although similar trends existed for malathion and methyl parathion, differences were not statistically significant. These results match previously published toxicity data where atrazine, although not acutely toxic even at much higher levels, decreased EC₅₀ values for chlorpyrifos by a magnitude of 4, decreased methyl parathion values by a magnitude of 2, and did not decrease values for malathion.     

Solid phase microextraction applied to the analysis of organophosphorus insecticides in fruits

Trace amounts of organophosphorus pesticides (OPs) were determined in various fruits by headspace solid phase microextraction (HS-SPME) and gas chromatography–nitrogen phosphorous detection (GC-NPD). Sampling from the headspace enhanced method selectivity, whereas at the same time improved fiber life time and method sensitivity. Diazinon, parathion, methyl parathion, malathion and fenithrothion were determined in various fruits: more than 150 samples of 21 types of fruits were studied. SPME-GC-NPD provided a useful and very efficient analytical tool: method linearity ranged from 1.2 to 700 ng/ml. Limits of detection (LODs) and quantitation (LOQs) ranged from 0.03 to 3 ng/ml and 0.12 to 10 ng/ml respectively, values well below the residue limits set by the EU. Less than 2% of the samples were found positive containing amounts higher than the EU limits. The effect of fruit peeling and washing was also investigated.     

Monoclonal antibody produced by heterologous indirect enzyme-linked immunosorbent assay and its application for parathion residue determination in agricultural and environmental samples

A heterologous indirect enzyme-linked immunosorbent assay (ELISA) was developed, which was based on monoclonal antibody (Mab) to determine parathion residue in agricultural and environmental samples. Eight Mabs were produced by introducing the heterologous indirect ELISA into the screening procedure. It was shown that these Mabs had more broad-reactivity with twenty competitors than that of 5H7 (Mab produced by homologous screening). So it became much easier using these new Mabs to develop heterologous immunoassays. In addition, all the developed heterologous ELISAs could be used to determine parathion residue in semiquantitative or quantitative level, and their detection limitation (LOD) was around 2 ng/mL. Compared with the previous heterologous ELISA (hapten 11/5H7) with IC₅₀ of 13.3 ng/mL, a better heterologous ELISA (hapten 11/1E1) was obtained with IC₅₀ of 3.8 ng/mL, which improved the sensitivity 3.5 times. Finally, the latter was applied to parathion residue determination in spiked agricultural and environmental samples without extraction or cleanup. The average recoveries of parathion added to water, soil, cucumber, Chinese cabbage and carrot were between 80.4 % and 111.8 %. The LOD for water and soil samples was 5 ng/mL, and the LOD for cucumber, rice and corn samples was 10 ng/mL.     

Heterogeneous reactions of suspended parathion, malathion, and fenthion particles with NO(3) radicals

Organophosphorus pesticides (OPPs) emit into the atmosphere in both gas and particulate phases via spray drift from treatments and post-application emission, but most of their degradations in the atmosphere are not well known. In this study, the heterogeneous reactions of nitrate (NO₃) radicals with three typical OPPs (parathion, malathion, and fenthion) absorbed on azelaic acid particles are investigated using an online vacuum ultraviolet photoionization aerosol time-of-flight mass spectrometer (VUV-ATOFMS). The reaction products observed with the VUV-ATOFMS are identified on the basis of GC/MS analysis of the products in the reaction between NO₃ radicals and the coating of the pesticide. Paraoxon is identified as the only product of parathion; malaoxon and bis(1,2-bis-ethoxycarbonylethyl)disulfide as the products of malathion; fenoxon, fenoxon sulfoxide, fenthion sulfoxide, fenoxon sulfone, and fenthion sulfone as the products of fenthion. The degradation rates of parathion, malathion, and fenthion under the experimental conditions are 5.5 × 10⁻³, 5.6 × 10⁻², and 3.3 × 10⁻² s⁻¹, respectively. The pathways of the heterogeneous reactions between the three OPPs and NO₃ radicals are proposed. The experimental results reveal the possible transformations of these OPPs through the oxidation of NO₃ radicals in the atmosphere.     

Simultaneous gas chromatographic determination of chlorpyrifos and its impurity sulfotep in liquid pesticide formulations

An analytical method for simultaneous determination of the active substance (chlorpyrifos) and its relevant impurity (sulfotep) in commercial pesticide formulations has been developed and validated. The proposed method entails extraction of the analytes from samples by sonication with acetone and analysis by gas chromatography-flame ionization detection (GC-FID). The proposed method was characterized by satisfactory accuracy and precision. The repeatability expressed as relative standard deviation (RSD) was lower than the acceptable values calculated from the modified Horwitz equation whereas individual recoveries were in the range of 98–102% and 80–120% for chlorpyrifos and sulfotep, respectively. The limit of quantification (LOQ) for the impurity (sulfotep) was 0.003 mg mL^?1 corresponding to the maximum permitted level according to Food and Agricultural Organization of the United Nations (FAO) specifications for the active substance (chlorpyrifos) being 3 g kg^?1 of the chlorpyrifos content found. The main advantage of the proposed method was a considerable reduction in the analysis time since both analytes were determined based on a single injection into the GC-FID. Analysis of real samples of commercial pesticide formulations confirmed fitness-for-purpose of the proposed method.     

Optimizing microwave-assisted solvent extraction (MASE) of pesticides from soil

Microwave-assisted solvent extraction (MASE) was investigated as an alternative for extraction of parathion (O,O-diethyl O-4-nitrophenyl phosphorothioate), methyl parathion (O,O-dimethyl O-4-nitrophenyl phosphorothioate), p,p'-DDE [1,1'-dichloro-2,2-bis(4-chlorophenyl)ethane], hexachlorobenzene (HCB), simazine (6-chloro-N2,N4-diethyl- 1 ,3,5-triazine-2,4-diamine) and paraquat dichoride (1,1'-dimethyl-4,4'-bipyridinium) from two different soils and from an earthworm-growing substrate. The matrices were fortified with 14C-radiolabeled pesticides and extracted with various solvent systems under different microwave conditions. Recoveries of more than 80% could be obtained depending on the used microwave conditions and solvent, except for paraquat whose recovery was generally less efficient. Thus, MASE can be successfully used to extract pesticides from environmental and biological samples and could be a viable alternative to conventional extraction methods. The technique uses smaller amounts of organic solvents, thereby minimizing the costs of the analysis and the disposal of waste solvent.     

Development of IC-ELISA for detection of organophosphorus pesticides in water

Diethyl (carboxymethyl) phosphonate (DECP) was used as the hapten to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for detecting organophosphorus pesticides (OPs). Conjugator of DECP with bovin serum albumin (BSA) was used as the immunogen for producing the polyclonal antibodies (PcAbs). Three antisera were obtained after the immune procedure. Characterization studies of the PcAbs indicated that the titer of antiserum-1 was highest in 3 antisera, and antiserum-1 had high affinity and specificity to the parathion, dichlorvos and pirimiphos. The IC-ELISA showed an IC₅₀ of 0.428 μ g/mL with a detection limit of 0.0125 μ g/mL to parathion. The assay also indicated that the IC₅₀ values of pirimiphos and dichlorvos were 0.331 μ g/mL and 1.25 μ g/mL respectively, and the detection limits of pirimiphos and dichlorvos were 0.0116 μ g/mL and 0.048 μ g/mL respectively. Recoveries of parathion, pirimiphos and dichlorvos spiked into water samples ranged from 90% to 160%. The results indicated that the ELISA could be a convenient and supplemental analytical tool for monitoring OPs residues in environmental water samples.     

Pesticide decontamination from fabric by laundering and simulated weathering1

Abstract

This laboratory study investigated the effectiveness of selected detergents and the phenomenon of simulated environmental conditions (weathering) on the removal of a commercial‐grade mixture of parathion and methyl parathion from a three‐layer laminated fabric. The weathering treatment consisted of exposure and non‐exposure to simulated environmental conditions of heat, light, and humidity. Contaminated fabric samples were laundered in one of three detergents containing an anionic, a nonionic, and a combined anionic and nonionic surfactant. The test fabric, a three‐layer fabric containing an impermeable microporous film laminated between two layers of nylon, was pipette‐contaminated with 400 microliters of field strength pesticide solution and allowed to dry. Half of the contaminated samples were weathered in an Atlas Fade‐Ometer. All of the contaminated samples were subsequently laundered in a Launder‐Ometer. Percent of pesticide residue was determined by gas chromatography. Weathering did significantly reduce both parathion and methyl parathion residues remaining in the test fabric. No statistically significant difference was found among the three detergents. High amounts of both parathion and methyl parathion remained in the test fabric after weathering and laundry treatments. Before the test fabric can be recommended for use in protective garments further research is needed to develop more effective decontamination procedures.     

Multiresidue determination of carbamate,organochlorine, organophosphorus,and dicarboximide pesticides in lettuce by GC/MS

An extraction method based on matrix solid-phase dispersion was developed to determine pirimicarb, methyl parathion, malathion, procymidone, α -endosulfan and β -endosulfan in lettuce using gas chromatography-mass spectrometry. The best results were obtained using 4.0 g of lettuce, 2.0 g of silica as dispersant sorbent, 0.1 g of activated carbon as clean up sorbent and acetonitrile as eluting solvent. The method was validated using lettuce samples fortified with pesticides at six different concentration levels (0.1 to 2.0 mg/kg). Average recoveries (7 replicates) ranged from 50 to 120 %, with relative standard deviations between 0.6 and 8.0 %. Detection and quantification limits for lettuce ranged from 0.01 to 0.02 mg/kg and 0.04 to 0.10 mg/kg, respectively.     

Pesticide decontamination from fabric by laundering and simulated weathering

This laboratory study investigated the effectiveness of selected detergents and the phenomenon of simulated environmental conditions (weathering) on the removal of a commercial-grade mixture of parathion and methyl parathion from a three-layer laminated fabric. The weathering treatment consisted of exposure and non-exposure to simulated environmental conditions of heat, light, and humidity. Contaminated fabric samples were laundered in one of three detergents containing an anionic, a nonionic, and a combined anionic and nonionic surfactant. The test fabric, a three-layer fabric containing an impermeable microporous film laminated between two layers of nylon, was pipette-contaminated with 400 microliters of field strength pesticide solution and allowed to dry. Half of the contaminated samples were weathered in an Atlas Fade-Ometer. All of the contaminated samples were subsequently laundered in a Launder-Ometer. Percent of pesticide residue was determined by gas chromatography. Weathering did significantly reduce both parathion and methyl parathion residues remaining in the test fabric. No statistically significant difference was found among the three detergents. High amounts of both parathion and methyl parathion remained in the test fabric after weathering and laundry treatments. Before the test fabric can be recommended for use in protective garments further research is needed to develop more effective decontamination procedures.     

Distribution of organophosphate insecticides in a Thai biomass-water system

Sorption and leaching of the organophosphate (OP) pesticides chlorpyrifos, profenofos, methyl parathion and malathion were investigated with four different types of biomass: coconut husk, rice husk, peat moss and peanut shell. Organic carbon contents of the biomass were in the range of 35.4–45.4%. Sorption studies were carried out by the batch (equilibrium) method at 4 different OP spike concentrations and at pH 3–7. Sorption isotherms conformed to a linear Freundlich equation and the Freundlich constant or sorption coefficient (K_D) confirmed that biomass organic carbon content was the principal sorbent factor affecting OP sorption. For a given sorbent, correlation of the extent of sorption with sorbate chemical properties was examined. Column leaching experiments involving different masses of coconut husk and peanut shell pre-spiked with OPs at the level equivalent to actual spraying concentrations in some Thai tangerine orchards were conducted. These experiments included repeated spikings and leaching. A water flow rate of 20 mL min^?1 was employed corresponding to the current average watering regime. Retardation and biodegradation with these sorbents were also examined.     

Determination of pesticide residues in IPM and non-IPM samples of mango (Mangifera indica)

Studies were conducted to analyze the residue of commonly used pesticides viz. methyl parathion, chloropyrifos, endosulfan, cypermethrin, fenvalerate, carbendazim, imidacloprid and carbaryl in mango, Dashehari variety, integrated pest management (IPM) and non-IPM samples were collected from the IPM and non-IPM orchards, Lucknow, India. We also present a method for the simultaneous determination of these pesticides in mango samples. Residues of methyl parathion, chloropyriphos, endosulfan, cypermethrin, fenvalerate were extracted from the samples with acetone: cyclohexane: ethyl acetate in the ratio 2:1:1 followed by cleanup using neutral alumina. Analysis was performed by gas chromatography-electron capture detector (GC-ECD) with a megabore column (OV-1). Residues of carbendazim, imidacloprid and carbaryl were extracted with acetone and after cleanup, analysis was performed by high performance liquid chromatography (HPLC) using photo diode array (PDA) detector. Recoveries of all the pesticides ranged between 72.7 – 110.6%, at 0.1 and 1.0 μg g^{? 1} level of fortification. The residues detected in non-IPM samples of mango were found to be below the prescribed limits of maximum residue limit (MRL) while IPM samples were free from pesticide residues.     

The rate of soil reduction as affected by levels of methyl parathion and 2, 4‐D

Abstract

A simple technique was demonstrated for determining the potential for synthetic organics to stress microbial populations. Oxidized Crowley and Cecil soil materials were amended with varying concentrations of 2,4‐D and methyl parathion, flooded, and then analyzed for changes in pH, redox potential, and levels of soluble plus exchangeable Fe, Mn, and Zn, all of which may be directly or indirectly influenced by the activity of soil microorganisms. At the concentrations tested (up to 75 ppm), there was little effect of 2,4‐D, but methyl parathion apparently did affect microbial activity contributing to changes in the measured soil properties upon flooding. This approach may be a useful technique for screening various compounds for their potential to stress microbial activity that, for many researchers, would be easier than direct observations of microbial parameters such as population numbers and classifications, and enzyme levels.     

Extraction procedures for the study of phytotoxicity and degradation processes of selected triketones in a water ecosystem

Simple and effective extraction methods based on matrix solid-phase dispersion (MSPD), dispersive liquid–liquid microextraction (DLLME), and solid-phase extraction (SPE) coupled with high-performance liquid chromatography with diode array detector (HPLC-DAD) were developed to determine triketone herbicides—sulcotrione (SUL), mesotrione (MES), tembotrione (TEMB), and their degradation products—in plant tissues and water samples. The extraction procedures were employed to enable quantification of the accumulation of selected triketone herbicides and their degradation products in a model aquatic plant, Egeria densa. To obtain comprehensive information about the triketones' influence on an aquatic plant, changes in chlorophyll concentration in plants exposed to these triketones were monitored. The average recovery ranged from 58 to 115 % (coefficients of variation 7–12 %) for plant tissues and from 52 to 96 % (coefficients of variation 8–20 %) for water samples. The limit of detection (LOD) for the MSPD–HPLC-DAD procedure was in the range of 0.06–0.23 μg/g, whereas for DLLME–HPLC-DAD and SPE–HPLC-DAD, LOD was in the range of 0.06–0.26 μg/mL. Symptoms of the phytotoxicity of sulcotrione, mesotrione, tembotrione, and their degradation products (decrease of chlorophyll concentration in plant sprouts) were observed for E. densa cultivated in water with herbicide concentrations of 100 μg/L. Moreover, the tembotrione degradation product exhibited a high level of accumulation and low metabolism in plant tissues in comparison to the other triketones and their degradation products.     

A rapid and sensitive fluoroimmunoassay based on quantum dot for the detection of chlorpyrifos residue in drinking water

A rapid and sensitive indirect competitive fluorescence-linked immunosorbent assay (cFLISA) method based on quantum dots as the fluorescence label coupled with secondary antibody (Ab₂) for the detection of chlorpyrifos in drinking water has been developed. The cFLISA method allowed for chlorpyrifos determination in a liner working range of 15.2–205.5 ng mL^?1. The 50 % inhibition value (IC₅₀) and the limit of detection (LOD) of the cFLISA were 50.2 ng mL^?1 and 8.4 ng mL^?1, while the IC₅₀ and the LOD of the conventhional enzyme linked immunosorbent assay (ELISA) were 95.3 ng- mL^?1 and 16.2 ng mL^?1, respectively. When the concentrations of chlorpyrifos were 200, 100 and 50 ng mL^?1, the recoveries ranged from 90.8 % to 108.2 % with a coefficient of variation (CV) of 7.5 %–15.2 %. In water sample analysis, the results of cFLISA were similar to those obtained from a cELISA and a high performance liquid chromatography (HPLC) method, while the detection time by cFLISA was reduced 0.5 h compared with ELISA. It showed that cFLISA could be used as a new screening method for the detection of pesticide residue.     

Rapid determination of residues of pesticides in honey by µGC-ECD and GC-MS/MS: Method validation and estimation of measurement uncertainty according to document No. SANCO/12571/2013

A simple and straightforward method for simultaneous determination of residues of 13 pesticides in honey samples (acrinathrin, bifenthrin, bromopropylate, cyhalothrin-lambda, cypermethrin, chlorfenvinphos, chlorpyrifos, coumaphos, deltamethrin, fluvalinate-tau, malathion, permethrin and tetradifon) from different pesticide classes has been developed and validated. The analytical method provides dissolution of honey in water and an extraction of pesticide residues by n-Hexane followed by clean-up on a Florisil SPE column. The extract was evaporated and taken up by a solution of an injection internal standard (I-IS), ethion, and finally analyzed by capillary gas chromatography with electron capture detection (GC-µECD). Identification for qualitative purpose was conducted by gas chromatography with triple quadrupole mass spectrometer (GC-MS/MS). A matrix-matched calibration curve was performed for quantitative purposes by plotting the area ratio (analyte/I-IS) against concentration using a GC-µECD instrument. According to document No. SANCO/12571/2013, the method was validated by testing the following parameters: linearity, matrix effect, specificity, precision, trueness (bias) and measurement uncertainty. The analytical process was validated analyzing blank honey samples spiked at levels equal to and greater than 0.010 mg/kg (limit of quantification). All parameters were satisfactorily compared with the values established by document No. SANCO/12571/2013. The analytical performance was verified by participating in eight multi-residue proficiency tests organized by BIPEA, obtaining satisfactory z-scores in all 70 determinations. Measurement uncertainty was estimated according to the top-down approaches described in Appendix C of the SANCO document using the within-laboratory reproducibility relative standard deviation combined with laboratory bias using the proficiency test data.     

Genotoxicity of methyl parathion in short‐term bacterial test systems

Abstract

Genotoxicity of the insecticide methyl parathion was investigated in Salmonella typhimurium and Escherichia coli bacterial test systems for the detection of back mutations and DNA‐damage. Methyl parathion was mutagenic to S. typhimurium strain TA100 after activation with rat liver microsomal and cytosolic enzymes. In DNA repair tests, methyl parathion was effective in inducing damage to the S. typhimurium strain TA1538 which lack excision repair compared to the strain TA1978 which is proficient in excision repair mechanisms. Normal laboratory light conditions had no effect on the mutagenicity tests, however, exposure of methyl parathion in the petri dish containing the tester strain TA100 and rat liver microsomal and cytosolic enzymes reduced the mutagenic activity and increased the toxic effects of methyl parathion.