Abstract
Diniconazole hapten was synthesized and conjugated to bovine serum albumin (BSA) by the carbodiimide method to produce an immunogen and to ovalbumin (OVA) by mixed anhydride method to produce a coating antigen. Polyclonal antibody against diniconazole was generated by immunizing New Zealand rabbits with the immunogen. Under optimised conditions (20% methanol, 0.4 mol/L Na +, pH 7.5), an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for detecting diniconazole. The 50% inhibitory concentration (IC 50) was 0.071 ± 0.013 mg/L and the limit of detection (LOD) was 1.28 ± 0.80 μg/L. Triazole fungicide analogues of diniconazole were tested and did not obviously cross-react, except for uniconazole (2.25%). The recoveries obtained after addition of standard diniconazole to agricultural samples, including water, soil, pear, grape, tomato and wheat flour, ranged from 70% to 120%. The ic-ELISA developed could successfully be applied to analysis of diniconazole residues in agriculture samples.
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