Development of competitive enzyme-linked immunosorbent assays for boscalid determination in fruit juices |
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Authors: | Antonio Abad-Fuentes,Francesc A. Esteve-Turrillas,Consuelo Agulló ,Antonio Abad-Somovilla,Josep V. Mercader |
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Affiliation: | 1. Department of Biotechnology, IATA-CSIC, Agustí Escardino 7, 46980 Paterna, València, Spain;2. Department of Organic Chemistry, Universitat de València, Doctor Moliner 50, 46100 Burjassot, València, Spain |
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Abstract: | Boscalid is a modern, broad-spectrum carboxamide pesticide highly efficient against most fungal diseases affecting valuable crops. In this study, a boscalid-mimicking derivative with a six-carbon spacer arm replacing the chlorine atom at the pyridine ring of the target molecule was synthesized and coupled to carrier proteins. Following rabbit immunization, antibodies against this agrochemical were obtained for the first time, and they were characterised in terms of affinity and specificity, tolerance to solvents, and robustness to changes in buffer pH and ionic strength, using two assay formats. Both of the optimised immunoassays showed limits of detection below 0.1 μg/L. Moreover, matrix effects of grape, peach, apple, and tomato juices were evaluated. Finally, a simple and easy procedure was set up for boscalid determination with spiked samples, affording limits of quantification of 10 μg/L, a value well below the sensitivity levels required for monitoring campaigns of pesticide residue analysis in food. |
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Keywords: | Pesticide Fungicide Hapten Antiserum Immunoassay Competitive ELISA Agrochemical residues Food safety |
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