Development of a sensitive competitive indirect ELISA for parathion residue in agricultural and environmental samples

A sensitive competitive indirect enzyme-linked immunosorbent assay (ELISA) for the insecticide parathion was developed. The optimal immunogen was 5-(ethoxy(4-nitrophenoxy)phosphorothioylamino)pentanoic acid. In addition, five competitors were applied for development of a heterologous competitive indirect ELISA. Then several physicochemical factors (organic solvent, ionic strength and pH) that influence assay performance were studied and optimized. The IC50 and IC10 of the optimized ELISA were 0.95 and 0.15 ng/mL, respectively, which meant almost 86-fold and 6-fold improvement in the assay sensitivity in comparison with the homologous assay (81.74 ng/mL) and the non-optimized heterologous assay (5.60 ng/mL). Finally, the assay was applied to the analysis of parathion in spiked agricultural and environmental samples without extraction or cleanup. The average recoveries of parathion added to water, soil, cucumber, rice and corn were between 78.57% and 107.67%. The limit of detection (LOD) for water and soil samples was 5 ng/mL, and the LOD for cucumber, rice and corn samples was 10 ng/mL.