Development of Two Enzyme-Linked Immunosorbent Assays for Detection of Endosulfan Residues in Agricultural Products

Two competitive immunoassays, a laboratory assay based on microwell plates and a field test based on the use of polystyrene tubes, have been developed for the detection of endosulfan in agricultural products. The limit of detection for the microwell plate format was 0.8 ± 0.1 μg/kg, and the limit of detection for the tube format was 1.6 ± 0.2 μg/kg. A simple, rapid, and efficient extraction method was employed, and 76−112% recoveries of spiked samples were obtained. Methanol extracts of some agricultural product samples such as grape, carrot, spinach, and tobacco could be analyzed directly by immunoassay after dilution in 0.5% fish skin gelatin−phosphate buffered saline. In contrast, extracts of green tea caused significant interference in the assay, and a number of simple cleanup methods were ineffective in removing interference. However, use of the coagulating reagent polyvinyl pyrrolidone removed the matrix effect effectively. For the validation of the enzyme-linked immunosorbent assay (ELISA) tests, samples were analyzed by ELISA and gas chromatography (GC) after solid phase extraction. The relationship between data obtained using the tube assay and microwell assay was good (the lowest r² value was 0.94), and also, the immunoassay assay data correlated well with data obtained from GC analysis (the lowest r² value was 0.93). The developed immunoassay methods are the suitable methods for the rapid quantitative and reliable determination of endosulfan residues in agricultural products.

Keywords: Immunoassay; endosulfan; agricultural products; matrix effect; validation