Czech J. Food Sci., 2016, 34(5):429-438 | DOI: 10.17221/275/2015-CJFS

Enzyme-linked immunosorbent assay for simultaneous detection of two fungicides kresoxim-methyl and trifloxystrobin in orangesFood Analysis, Food Quality and Nutrition

Wei Yang, Jie Zhu, Ming-Qi Liu, Xian-Jun Dai
College of Life Sciences, China Jiliang University, Hangzhou, P.R. China

To assemble an indirect competitive enzyme-linked immunosorbent assay (ELISA) for estimating two strobilurin fungicides at the same time, the hapten was synthesised which contained the common active group, (E)-2-(2-bromo-phenyl)-2-(methoxyimino) acetic acid methyl ester (OEBr) in kresoxim-methyl and trifloxystrobin. The immunogen and coating antigen were respectively prepared through conjugating the above-mentioned hapten with BSA and OVA by the mixed anhydride and activated ester methods, and polyclonal antibodies were produced by immunised rabbits. An enzyme-linked immunosorbent assay was developed for simultaneous detection of kresoxim-methyl and trifloxystrobin. In ELISA, the antiserum showed high affinity and sensitivity to kresoxim-methyl and trifloxystrobin, and their IC50 value and detection limit (expressed as IC10) were 14.7 and 0.0044 µg/ml, respectively, for kresoxim-methyl, and 22.9 and 0.014 µg/ml, respectively for trifloxystrobin. The cross-reaction rate was below 0.1% for other strobilurin fungicides. Recovery study of ELISA from spiked samples of homogenised peeled oranges (final concentrations of 100, 10, and 1 µg/ml) resulted in recovery levels in the range of 82-104%.

Keywords: kresoxim-methyl; trifloxystrobin; polyclonal antibody; ELISA; recovery rate

Published: October 31, 2016  Show citation

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Yang W, Zhu J, Liu M, Dai X. Enzyme-linked immunosorbent assay for simultaneous detection of two fungicides kresoxim-methyl and trifloxystrobin in oranges. Czech J. Food Sci. 2016;34(5):429-438. doi: 10.17221/275/2015-CJFS.
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