Competitive enzyme-linked immunosorbent assay for the determination of the phenylurea herbicide chlortoluron in water and biological fluids

Abstract

A competitive ELISA method suitable for the monitoring of the herbicide chlortoluron [N-(3-chloro-4-methylphenyl)-N′-dimethylurea] in different types of water and biological fluids was developed. The production of the immunogen utilized in this work was achieved by covalently coupling bovine thyroglobulin with the synthesized hapten (N′-3-chloro-4-methylphenyl-N-carboxypropyl urea) using the N-hydroxysuccinimide active ester method. The chlortoluron antibody, raised in sheep after immunization with the immunogen, showed no cross-reactivity with a large range of pesticides, although some cross-reactivity was displayed with various phenylurea herbicides (i.e., chlorbromuron, isoproturon and metoxuron). The limit of detection of the chlortoluron ELISA method was 0.015 µg l^–1, well below the legal European limit for individual pesticides in drinking water (the EC maximum admissible concentration, 0.1 µg l^–1). In addition, reproducible and quantitative recovery of chlortoluron from water, obtained from various sources, and biological fluids was possible without any sample preparation. The ELISA technique for chlortoluron developed and described here proved to be rapid, sensitive and specific, fulfilling the needs of present legislation relating to the use and levels of pesticides in the environment.