Development of an indirect competitive enzyme-linked immunosorbent assay for detecting flunixin and 5-hydroxyflunixin residues in bovine muscle and milk
Development of an indirect competitive enzyme-linked immunosorbent assay for detecting flunixin and 5-hydroxyflunixin residues in bovine muscle and milk
复制标题
开发用于检测牛肌肉和牛奶中氟尼辛和5-羟基氟尼辛残留的间接竞争酶联免疫吸附测定法
DOI：
10.1080/09540105.2019.1577365
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发表时间：
2019-01
影响因子：
3
通讯作者：
Jiang Wenxiao
中科院分区：
文献类型：
--
作者：
Chen Xiaona;Peng Shimin;Liu Chen;Zou Xin;Ke Yuebin;Jiang Wenxiao
摘要 动物源性食品中氟尼辛残留可能对人类健康造成危害。本文以5-羟基氟尼辛偶联牛血清白蛋白为免疫原，获得了广泛选择性的多克隆抗体。多克隆抗体的半抑菌浓度（IC50），氟尼辛为1.43ng/mL，5-羟基氟尼辛为0.29ng/mL，远低于美国、欧盟和中国规定的最大残留限量。牛肌肉中氟尼辛的检出限为2.98μg/kg，牛奶中5-羟基氟尼辛的检出限为0.78μg/L。在加标和回收率测试中，平均回收率范围为 83% 至 105%，方差系数范围为 5.8% 至 11.3%，令人满意。此外，ELISA 方法还通过成熟的 LC-MS/MS 方法进行了验证。结果表明，所建立的方法适用于氟尼辛和5-羟基氟尼辛残留量的分析。
ABSTRACT The flunixin residues in foods of animal origin could pose hazards for human health. In this paper, a broad-selectivity polyclonal antibody was obtained using 5-hydroxyflunixin coupled with bovine serum albumin as the immunogen. The semi-inhibitory concentration (IC50) of the polyclonal antibody was 1.43 ng/mL for flunixin and 0.29 ng/mL for 5-hydroxyflunixin, respectively, which were far below the maximum residue limits set by the United States, the European Union and China. The limits of detection were calculated to be 2.98 μg/kg for flunixin in bovine muscle and 0.78 μg/L for 5-hydroxyflunixin in milk. In spike and recovery tests, the average recovery ranged from 83% to 105%, with a satisfying coefficient of variance ranging from 5.8% to 11.3%. Furthermore, the ELISA method was validated by a well-established LC-MS/MS method. These results proved that the established method is suitable for flunixin and 5-hydroxyflunixin residue analysis.
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