Tube-Immunoassay for Rapid Detection of Carbaryl Residues in Agricultural Products

An antibody-based rapid, quantitative, and qualitative tube enzyme-linked immunosorbent assay (tube-ELISA) was developed and used to determine carbaryl (1-naphthyl methylcarbamate) residues in agricultural products (apple, Chinese cabbage, rice, and barley). The tube-ELISA is a competitive immunoassay in which the antibody is coated in the polystyrene tube, with a dynamic range between 0.7 and 46.3 μg kg^?1. Carbaryl was extracted from each agricultural sample by hand-shaking with methanol and examined for application to on-site analysis. After the liquid extraction, the sample extracts diluted with buffer were analyzed by rapid tube-ELISA directly. The overall test time was around 15–30 min, including sample preparation and assay performance. The results obtained from tube-ELISA correlated well with high-performance liquid chromatography (R ² > 0.9). The study shows that tube-ELISA is useful as a quality control tool and can be used to quantitatively detect carbaryl as well.     

Rapid enzyme immunoassays for the detection of carbaryl and methoprene in grains

In order for grain handlers and traders to reliably estimate residues of grain protectants in the field, antibody-based rapid tests were developed for carbaryl (1-naphthyl methylcarbamate) and methoprene [isopropyl (E,E)-(RS)-11-methoxy-3,7,11-trimethyldodeca-2,4-dienoate]. To complement the rapid analysis, a simple and rapid extraction technique was developed. In these tests, a pesticide-containing methanol extract of the grain sample and an enzyme-labeled component are added to precoated strips. After a brief incubation, the strips are washed and a substrate/chromogen for the enzyme is added. The color developed is stopped by acidification and the results are read either by eye or in a portable field photometer. The overall test time is under 20 minutes. For carbaryl, the test had a limit of detection of 4.5 ppb (1.1 ppm in grain), while the methoprene test had a limit of detection of 4 ppb (1 ppm in grain) based on the lower datum point, which is 15% inhibition, in the standard curves. Both assays can be used as a screening test for carbaryl and methoprene in animal feed grains.     

RAPID ENZYME IMMUNOASSAYS FOR THE DETECTION OF CARBARYL AND METHOPRENE IN GRAINS

In order for grain handlers and traders to reliably estimate residues of grain protectants in the field, antibody-based rapid tests were developed for carbaryl (1-naphthyl methylcarbamate) and methoprene [isopropyl (E,E)-(RS)-11-methoxy-3,7,11-trimethyldodeca-2,4-dienoate]. To complement the rapid analysis, a simple and rapid extraction technique was developed. In these tests, a pesticide-containing methanol extract of the grain sample and an enzyme-labeled component are added to precoated strips. After a brief incubation, the strips are washed and a substrate/chromogen for the enzyme is added. The color developed is stopped by acidification and the results are read either by eye or in a portable field photometer. The overall test time is under 20 minutes. For carbaryl, the test had a limit of detection of 4.5 ppb (1.1 ppm in grain), while the methoprene test had a limit of detection of 4 ppb (1 ppm in grain) based on the lower datum point, which is 15% inhibition, in the standard curves. Both assays can be used as a screening test for carbaryl and methoprene in animal feed grains.     

Reductive degradation of carbaryl in water by zero-valent iron

Reduction of carbaryl solution by zero-valent iron powder (ZVIP) was studied in a rotator batch system (70 rpm) in order to evaluate the utility of this reaction in remediation of carbamate contaminated water. Degradation with different amount of ZVIP: 0.01, 0.02, 0.03, 0.04 g/ml at pH 6.6 and at ambient temperature was investigated. The results show that the process exhibits a degradation rate appearing to be directly proportional to the surface contact area of ZVIP (325-mesh) with the carbaryl molecules. Three analytical techniques were used to monitor carbaryl degradation: (1) A UV-Vis diode array spectrophotometer was used to record all spectra. (2) A high performance liquid chromatography was used to separate by-products and examine the evolution of breakdown products. (3) A home-built spectrophosphorimeter that uses the solid surface room temperature phosphorescence (SSRTP) was employed to observe selectively the decline of the carbaryl concentration at different amount of ZVIP on Whatman no. 1 filter paper. Results show that the reducing degradation of carbaryl with ZVIP as the source of electrons is effective with a half-life close to several minutes.     

Dissipation of carbofuran and carbaryl on Oolong tea during tea bushes, manufacturing and roasting processes

Carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranol-N-methylcarbamate) and carbaryl (1-naphthyl-N-methylcarbamate) are insecticides widely used in tea plantations. The objective of the present study was to evaluate the dissipation of carbofuran and carbaryl during the growth periods of Oolong tea, processing and roasting. Analysis of the residual insecticides was carried out using high-pressure liquid chromatography with a post-column fluorescence detector. Results showed that in the tea field carbofuran dissipated faster then carbaryl. Manufacturing processes of Oolong tea further reduced the carbofuran and carbaryl contents. The persistence of carbofuran and carbaryl was decreased with increasing roasting temperature. From the results, we conclude that the presence of carbofuran and carbaryl in tea can be reduced by proper field management, manufacturing and roasting processes.     

Dissipation of carbofuran and carbaryl on Oolong tea during tea bushes,manufacturing and roasting processes

Carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranol-N-methylcarbamate) and carbaryl (1-naphthyl-N-methylcarbamate) are insecticides widely used in tea plantations. The objective of the present study was to evaluate the dissipation of carbofuran and carbaryl during the growth periods of Oolong tea, processing and roasting. Analysis of the residual insecticides was carried out using high-pressure liquid chromatography with a post-column fluorescence detector. Results showed that in the tea field carbofuran dissipated faster then carbaryl. Manufacturing processes of Oolong tea further reduced the carbofuran and carbaryl contents. The persistence of carbofuran and carbaryl was decreased with increasing roasting temperature. From the results, we conclude that the presence of carbofuran and carbaryl in tea can be reduced by proper field management, manufacturing and roasting processes.     

Removal of pesticides from rinsate by adsorption using agricultural residuals as medium

Abstract

The feasibility of using agricultural residual with highly organic matter content ‐ spent mushroom compost (SMC) as adsorption medium for removal of pesticides including carbaryl, carbofuran, and aldicarb with a concentration range of 0–30 mg/L from rinsate was investigated. Bottle‐point method was used to determine adsorption isotherms for single‐pesticide and multi‐pesticides. It was found that SMC might potentially be used for on‐farm treatment of pesticide rinsate. The adsorption capacity of SMC was increased compared to most soils, possibly related to the increased organic matter content. The rapid equilibrium of pesticide solution and SMC was consistent with a physical type of adsorption mechanism. The adsorption of carbamate pesticides on SMC was found to exhibit nonlinear “favorable”; adsorption behavior that could be characterized well by the Freundlich isotherm model. In addition, the multi‐pesticide adsorption tests displayed the characteristics of competitive behavior. The competitive ability of these three pesticides in multi‐components adsorption was in the order carbaryl > carbofuran > aldicarb, which was consistent with the adsorbability of pesticides.     

Predictability by Box-Behnken model for carbaryl adsorption by soils of Indian origin

This study was undertaken to investigate the adsorption capacity of carbaryl on four Indian soils with different physiochemical properties. A batch adsorption study was carried out in order to evaluate the maximum adsorption capacity of carbaryl using a Response Surface Methodology (RSM). The effects of operating parameter such as initial carbaryl concentration (1–20 mgL^?1), adsorbent dosage (0.5–6 g) and contact time (10–180 min) were examined. The proposed quadratic model for Box-Behnken design fits very well to the experimental data because it may be used to navigate design space according to ANOVA results. The regression co-efficient (R²) of the models developed and the results of validation experiments conducted at optimal conditions strongly suggests that the predicted values are in good agreement with experimental results. Contour and response surface plots are used to determine the interactions effects of main factors and optimal conditions of the process. The experiment can be utilized as a guideline for better understanding of carbaryl adsorption onto soil under different operating conditions. The results show that the forest soil is most efficient in binding carbaryl (Sevin) than the other types of soil tested.     

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 microg 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.     

Allium cepa derived EROD as a potential biomarker for the presence of certain pesticides in water

Allium cepa root length inhibition test is a well recommended bioassay for the evaluation of the toxicity of various polluted waters. The utility of EROD (7-ethoxy resorufin O-deethylase) as a potential biomarker of pesticide pollution was investigated using the Allium cepa system. Onion bulbs exposed to model water samples containing any of the six pesticides viz. 2,4-D, HCB, malathion, carbaryl, DDT and endosulphan were analyzed for EROD activity. The pesticide treatment resulted in the enhanced activity of the enzyme, with carbaryl and HCB causing 63- and 53-fold induction respectively with respect to the control at a dose of 1.2 ppb. The industrial wastewater samples from Ghaziabad city of Northern India resulted in about a 68-fold rise in the EROD activity, whereas the Aligarh samples did not exhibit any change within the statistical limit. These results suggest the presence of the test pesticides in the Ghaziabad sample and their absence in the Aligarh sample. Pesticide analysis in the test water samples by HPLC supported this to a large extent. Presence of cycloheximide in the test system brought down the EROD activity, equal to that of control, suggesting the de novo synthesis of the enzyme following the exposure of Allium cepa to pesticides. These studies suggest that the Allium cepa derived EROD can act as a potential biomarker of certain pesticides since even 1ppb of total/individual pesticides brought about >10-fold induction of EROD. We recommend the assay of EROD in the Allium cepa system as a presumptive test for the detection of these pesticides before using analytical techniques like HPLC.     

Development of a simple, accurate SPME-based method for assay of VOCs in column breakthrough experiments

Solid-phase microextraction (SPME) with gas chromatography is to be used for assay of effluent liquid samples from soil column experiments associated with VOC fate/transport studies. One goal of the fate/transport studies is to develop accurate, highly reproducible column breakthrough curves for 1,2-cis-dichloroethylene (cis-DCE) and trichloroethylene (TCE) to better understand interactions with selected natural solid phases. For SPME, the influences of the sample equilibration time, extraction temperature and the ratio of volume of sample bottle to that of the liquid sample (V(T)/V(w)) are the critical factors that could influence accuracy and precision of the measured results. Equilibrium between the gas phase and liquid phase was attained after 200 min of equilibration time. The temperature must be carefully controlled due to variation of both the Henry's constant (K(h)) and the fibre/gas phase distribution coefficient (K(fg)). K(h) decreases with decreasing temperature while K(fg) increases. Low V(T)/V(w) yields better sensitivity but results in analyte losses and negative bias of the resultant assay. High V(T)/V(w) ratio yields reduced sensitivity but analyte losses were found to be minimal, leading to better accuracy and reproducibility. A fast SPME method was achieved, 5 min for SPME extraction and 3.10 min for GC analysis. A linear calibration function in the gas phase was developed to analyse the breakthrough curve data, linear between a range of 0.9-236 microgl(-1), and a detection limit lower than 5 microgl(-1).     

Effects of carbaryl,chlorpyrifos and endosulfan on growth,reproduction and respiration of tropical epigeic earthworm,Perionyx excavatus (Perrier)

Effects of sub-lethal doses of carbaryl (1-Naphthyl-methylcarbamate), chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridinyl-phosphorothioate) and endosulfan (6,7,8,9,10,10-Hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin-3-oxide), respectively a carbamate, an organophosphate and an organochlorine insecticide on growth, reproduction and respiration of the tropical earthworm, Perionyx excavatus (Perrier) were investigated under laboratory conditions. The results showed significant reduction in biomass, production and hatching of cocoon and production of juveniles of the worms exposed to 0.75 to 3.03 mg/kg soil of carbaryl, 0.91 to 3.65 mg/kg soil of chlorpyrifos and 3.75 to 15.0 μg/kg soil of endosulfan corresponding to 12.5 to 50 % of LC₅₀ value of the respective insecticide for P. excavatus. Endosulfan was found most dangerous among the three insecticides followed by carbaryl and chlorpyrifos. There was no hatching of the worms at endosulfan treatment 5.0 μg/kg soil (25 % LC₅₀) or above while the highest dose of carbaryl and chlorpyrifos (50 % of LC₅₀) rendered respectively 87.13 and 24.84 % reductions in hatching as compared to control. Chlorpyrifos produced no change in respiration of the worms except at the highest dose, while the worms showed an increase in evolution of CO₂ at all doses of carbaryl and endosulfan. Based on the recommended agricultural dose of each insecticide, it was concluded that application of endosulfan and carbaryl was potentially dangerous to earthworms.     

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.     

Effects of carbaryl, chlorpyrifos and endosulfan on growth, reproduction and respiration of tropical epigeic earthworm, Perionyx excavatus (Perrier)

Effects of sub-lethal doses of carbaryl (1-Naphthyl-methylcarbamate), chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridinyl-phosphorothioate) and endosulfan (6,7,8,9,10,10-Hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin-3-oxide), respectively a carbamate, an organophosphate and an organochlorine insecticide on growth, reproduction and respiration of the tropical earthworm, Perionyx excavatus (Perrier) were investigated under laboratory conditions. The results showed significant reduction in biomass, production and hatching of cocoon and production of juveniles of the worms exposed to 0.75 to 3.03 mg/kg soil of carbaryl, 0.91 to 3.65 mg/kg soil of chlorpyrifos and 3.75 to 15.0 μg/kg soil of endosulfan corresponding to 12.5 to 50 % of LC(50) value of the respective insecticide for P. excavatus. Endosulfan was found most dangerous among the three insecticides followed by carbaryl and chlorpyrifos. There was no hatching of the worms at endosulfan treatment 5.0 μg/kg soil (25 % LC(50)) or above while the highest dose of carbaryl and chlorpyrifos (50 % of LC(50)) rendered respectively 87.13 and 24.84 % reductions in hatching as compared to control. Chlorpyrifos produced no change in respiration of the worms except at the highest dose, while the worms showed an increase in evolution of CO(2) at all doses of carbaryl and endosulfan. Based on the recommended agricultural dose of each insecticide, it was concluded that application of endosulfan and carbaryl was potentially dangerous to earthworms.     

Persistence and biodegradation of carbaryl in soils

Abstract

The persistence of the methylcarbamate pesticide carbaryl was studied in four soils under flooded conditions. A substantial portion of the pesticide was recovered from all soils even after 15 days of its application, with the recovery ranging from 37% in an alluvial soil to 73% in an acid sulfate soil. The degradation of carbaryl was more rapid under flooded conditions than under nonflooded conditions. A bacterium, Pseudomonas cepacia, isolated from a flooded soil amended with a related methylcarbamate pesticide carbofuran, degraded carbaryl in a mineral medium supplemented with yeast extract.     

Development of an efficient extraction method for oxytetracycline in animal manure for high performance liquid chromatography analysis

Oxytetracycline (2-(amino-hydroxy-methylidene)-4-dimethylamino-5,6,10,11,12a-pentahydroxy-6-methyl-4,4a,5,5a-tetrahydrotetracene-1,3,12-trione) is a major member of the tetracycline antibiotics family of which are widely administered to animals in concentrated animal feeding operations for purposes of therapeutical treatment and health protection. With the disposal of animal manure as fertilizer into agricultural land, tetracyclines enter the environment. However, tetracyclines chelate with multivalent cations and proteins, resulting in low extraction efficiencies from animal manure for tetracycline residue analysis. In this study an efficient extraction method for oxytetracycline from steer manure using methanol/water solution amended with chelating organic acid was developed for the analysis of high performance liquid chromatography. The effect of species and amount of amendment acids, shaking time, methanol/water ratio, manure weight, and repeated times of extraction was investigated. It was optimized to amend 2.5 g citric acid and 1.1 g oxalic acid with 10.0 g manure sample in a 50-ml centrifuge tube and extract with 15 ml methanol/water (9:1 in volume) by vigorously shaking for 30 min in a reciprocating shaker. After centrifugation at 11,000 rpm, supernatant is collected. Sample was extracted for a total of 3 times. The developed extraction method was further applied to extract oxytetracycline from fresh and aged cow manure, swine and poultry manure, and soil. Satisfactory recoveries ranging from (84.1 ± 2.4) % to (102.0 ± 3.1) % were obtained, demonstrating that the optimized extraction method is robust for oxytetracycline from different manure sample matrixes.     

Effect of carbaryl (carbamate insecticide) on acetylcholinesterase activity of two strains of Daphnia magna (Crustacea,Cladocera)

The present study was designed to investigate the effect of carbaryl (carbamate insecticide) on the acetylcholinesterase activity in two strains (same clone A) of the crustacean cladoceran Daphnia magna. Four carbaryl concentrations (0.4, 0.9, 1.8 and 3.7 µg L^?1) were compared against control AChE activity. Our results showed that after 48 h of carbaryl exposure, all treatments induced a significant decrease of AChE activities whatever the two considered strains. However, different responses were registered in terms of lowest observed effect concentrations (LOEC: 0.4 µg L^?1 for strain 1 and 0.9 µg L^?1 for strains 2) revealing differences in sensitivity among the two tested strains of D. magna. These results suggest that after carbaryl exposure, the AChE activity responses can be also used as a biomarker of susceptibility. Moreover, our results show that strain1 is less sensitive than strain 2 in terms of IC50-48 h of AChE activity. Comparing the EC50-48 h of standard ecotoxicity test and IC50-48 h of AChE inhibition, there is the same order of sensitivity with both strains.     

Novel pipette-tip graphene/poly (vinyl alcohol) cryogel composite extractor for the analysis of carbofuran and carbaryl in water

A novel pipette-tip extractor of a graphene/poly (vinyl alcohol) cryogel (graphene/PVA) composite sorbent was prepared to preconcentrate carbamate pesticides in environmental water samples before analysis with a gas chromatograph-flame ionization detector (GC-FID). This novel pipette-tip extractor with the graphene/PVA sorbent exhibited a high porosity when observed through a scanning electron micrograph (SEM). Under optimal conditions, using only 1.0 mL of sample and 0.75 mL of eluting solvent, the developed method provided a wide linear range of 10–700 ng mL^?1 and 10–500 ng mL^?1 with limit of detection (LOD) of 6.40 ± 0.18 and 9.17 ± 0.34 ng mL^?1 for carbofuran (2,3-dihydro-2,2-dimethylbenzofuran-7-yl methylcarbamate) and carbaryl (1-naphthyl methylcarbamate), respectively. The pipette-tip extractor provided high extraction efficiency with high accuracy indicated, by good recoveries in the range of 74.5 ± 4.8% to 119.7 ± 1.6% and 76 ± 15% to 114 ± 19% for carbofuran and carbaryl, respectively. In addition, the fabrication procedure showed a good pipette-tip extractor-to-pipette-tip extractor reproducibility with a relative standard deviation of 1.3–9.8% (n = 5). When the developed pipette-tip extractor was applied for the extraction of carbofuran and carbaryl in surface water samples near vegetable plantation areas, 25.9 ± 8.2 ng mL^?1 of carbofuran was found, and carbaryl was also detected in concentrations that ranged from 45.0 ± 4.0 to 191 ± 13 ng mL^?1.     

The excretion of pesticides in saliva and its value in assessing exposure

The possibility that pesticides are secreted in the saliva and that this secretion can be used to assess exposure to pesticides is discussed. Both Kepone and carbaryl were tested through a series of in vitro studies and in vivo experiments including both acute and subchronic studies. In the in vivo studies, both Kepone and carbaryl were detected in the saliva in the first twenty-four hours, but seldom thereafter. Analysis of urinary samples indicated a rapid clearance of the substances from the animals. In vitro uptake and efflux studies suggested that movement of both substances was by passive diffusion and reached a plateau after 60 minutes of incubation. Efflux rates were unaffected by experimental conditions, but were ten times faster for carbaryl.     

Effects of long- and short-term exposure to carbaryl on survival, growth and reproduction of Daphnia ambigua

In laboratory experiments a cladoceran, Daphnia ambigua, was exposed for six instars to the insecticide carbaryl at five different concentrations (1, 2, 3, 4, 5 microg litre(-1)) or received short-term (10-h) exposure to 5 microg carbaryl litre(-1) at different life stages of the animal. In the long-term exposure experiments carbaryl at 1 microg litre(-1) did not have any detectable effects on the animals, whereas 2 microg litre(-1) reduced the survival, growth and reproduction of the animals, and higher concentrations killed all the animals examined before they grew to the fifth instar. These results indicate that the range of sublethal concentrations of carbaryl for D. ambigua is narrow. In the short-term exposure experiments the animals did not suffer any damage when they were exposed to the chemical during the egg stage. However, their growth and egg production were reduced when the short-term exposure was conducted during the life stages from the final embryo stage to the third instar. The effect was most marked when the animals were treated during the first instar, indicating that this is the stage when the animals are most sensitive to the chemical.