Enzyme-linked immunosorbent assays for the insecticide fenitrothion: Influence of hapten conformation and sample matrix on assay performance

Abstract

This study aimed at developing competitive enzyme-linked immunosorbent assays (ELISAs) for the organophosphorus (OP) insecticide fenitrothion using a monoclonal antibody. The hapten used to obtain the antibody had an ideal structural feature that allowed minimal functional group sacrifice. By using the antibody and a coating antigen, a competitive indirect ELISA was developed, which showed an IC 50 of 14 ng mL −1 with a detection limit of 3.0 ng mL −1. A competitive direct ELISA using an enzyme tracer was also developed, which showed an IC 50 of 17 ng mL −1 with a detection limit of 1.6 ng mL −1. The antibodies in both assays showed negligible cross-reactivity with the metabolites of fenitrothion and other OP pesticides except with the insecticides parathion-methyl and parathion-ethyl. Recoveries of fenitrothion from fortified rice and lettuce samples were determined and the bias in the recovery values was rationalized by using the standard curves obtained in the matrix extract.