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Development and Evaluation of an Enzyme-Linked Immunosorbent Assay for the Determination of Thiacloprid in Agricultural Samples
Authors:Zhenjiang Liu  Ming Li  Haiyan Shi  Minghua Wang
Affiliation:1. Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University, Jiangsu Key Laboratory of Pesticide Science, Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing, 210095, China
Abstract:An enzyme-linked immunosorbent assay (ELISA) was developed based on a polyclonal antibody for the analysis of thiacloprid in agricultural samples. Thiacloprid hapten was synthesized and conjugated to bovine serum albumin to produce an immunogen and ovalbumin to produce a coating antigen. Polyclonal antibodies were obtained from immunized New Zealand white rabbits. Under optimal conditions (5 % methanol, 0.1 mol/L Na+, pH 5.5), the ELISA showed a 50 % inhibitory concentration (IC50) value of 0.01 mg/L and a limit of detection (IC10) of 0.47 μg/L. No obvious cross-reaction with the other structural analogs of neonicotinoid insecticides showed that the polyclonal antibodies had a high specificity for thiacloprid. The average recoveries from spiked water, soil, pear, and tomato were in the range of 80 to 119 %. The results of the ELISA were confirmed by high-performance liquid chromatography, and the correlation of the results from the two methods had a high correlation coefficient of 0.99 (n?=?3) in spiked samples (soil, pear, and tomato). The proposed ELISA could successfully be applied to the determination of thiacloprid residues in agricultural samples.
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