Development of an enzyme-linked immunosorbent assay for determination of the miticide bromopropylate

This paper reports for the first time the development of an immunoassay for the analysis of the miticide bromopropylate (BP). The chemical structure of the immunizing haptens was designed to maximize the recognition of the bis-bromophenyl group of BP. Thus, the assay uses polyclonal antibodies raised against 2,2-bis(4-bromophenyl)-N-2-hydroxyacetamide-butanoic acid (hapten 2) conjugated to keyhole limpet hemocyanin from horseshoe crab. A heterologous indirect competitive enzyme-linked immunosorbent assay (ELISA) has been developed that can detect BP down to 0.14 microg L(-1). The assay has been proven to tolerate a wide range of ionic strengths and pH values. Studies on the selectivity of this immunoassay have demonstrated a high recognition of related pesticides that contain a bis-halophenyl group in their structure. Other pesticides do not interfere in the analysis of BP using this immunochemical technique. Preliminary experiments have shown that BP can be directly analyzed in white wine samples down to 0.16 microg L(-1) without the necessity of a cleanup procedure prior to the ELISA.