An indirectly competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the quantitative detection of fenvalerate. The hapten FVa (α-carboxy-3-phenoxyphenyl-2-(4-chloro-phenyl)-3-methyl-butyrate) and FVb (N-2-(carboxybutyl)carbamoyl-3-phenoxyphenyl-2-(4-chlorophenyl)-3-methylbutyrate) for the fenvalerate were prepared starting from 2-(4-chlorophenyl)-3-methyl-butyric acid and α-cyano-3-phenoxyhenzyl-benzyl alcohol. The hapten FVa was conjugated to ovalbumin (OVA) to form the coating antigen (FVa-OVA); the hapten FVb was conjugated to bovine serum albumin (BSA) to form the immunogen (FVb-BSA). The rabbits were immunised by conjugate of FVb-BSA, and titres of anti-fenvalerate serum (1.28×104) was determined by non-competitive indirect ELISA. After optimisation of the ic-ELISA, such as pH values, ionic strengths, concentrations of methanol, pH 7.5, phosphate-buffered saline of 0.3 M Na+ and 30% methanol were determined to optimum assay condition. The medium inhibitory concentration (IC50) for fenvalerate was (1.19±0.09) mg/L, and limit of detection (LOD) was (0.017±0.002) mg/L. Pyrethroids, such as fenpropathrin, cyhalothrin, permethrin, cypermethrin, deltamethrin and bifenthrin did not cross-react significantly in this assay.
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enzyme-linked immunosorbent assay;
Document Type: Research Article
Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University, Nanjing, China
June 1, 2009
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