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A sensitive and specific ELISA for determining a residue marker of three quinoxaline antibiotics in swine liver

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Abstract

Methyl-3-quinoxaline-2-carboxylic acid (MQCA) is a possible residue marker for three quinoxaline veterinary medicines (olaquindox, mequindox, and quinocetone). The wide application of mequindox/quinocetone or the illegal use of olaquindox leads to MQCA residue in animal’s original food, thereby threatening the safety of human food. The indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) with a specific coating antigen and monoclonal antibody (MAB) was established and optimized for detecting MQCA in swine liver. Samples were acidified with 2 mol l−1 hydrochloric acid, extracted with ethyl acetate–hexane–isopropanol (8 + 1 + 1, v/v/v) and then detected by IC-ELISA. The logarithm correlation of standards to OD values ranged from 0.2 to 200 μg l−1, with IC50 of 6.46 μg l−1. Negligible cross-reactivity happened to five quinoxaline antibiotics (olaquindox, mequindox, quinocetone, carbadox, and cyadox) and the metabolite of carbadox and cyadox (quinoxaline-2-carboxylic acid). When spiked with 1 to 100 μg kg−1 of MQCA, the recoveries ranged from 85.44 to 100.02 %, with the intra-assay coefficient of variation (CV) of 6.64–10.57 % and inter-assay CV of 7.29–10.88 %. The limit of detection for MQCA was 1.0 μg kg−1 in swine liver. Furthermore, incurred samples were detected by the IC-ELISA and then conformed by a reported LC/MS/MS method, it shown that there was good correlation between the two methods. All these results indicated that the IC-ELISA method is appropriate for surveillance MQCA residue in animal tissues.

Synthesis route of 2-acrylic-1,4-binitrogen-quinoline combined to BSA(OVA) by active ester method

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Acknowledgment

This research was supported by the National Basic Research Program of China (No. 2009CB118801).

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Authors and Affiliations

  1. National Reference Laboratory for Veterinary Drug Residue Beijing Center, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China

    Linli Cheng, Jianzhong Shen, Zhanhui Wang, Wenxiao Jiang & Suxia Zhang

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  1. Linli Cheng
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  2. Jianzhong Shen
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  3. Zhanhui Wang
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  4. Wenxiao Jiang
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  5. Suxia Zhang
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Correspondence to Suxia Zhang.

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Cheng, L., Shen, J., Wang, Z. et al. A sensitive and specific ELISA for determining a residue marker of three quinoxaline antibiotics in swine liver. Anal Bioanal Chem 405, 2653–2659 (2013). https://doi.org/10.1007/s00216-012-6696-x

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  • DOI: https://doi.org/10.1007/s00216-012-6696-x

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