呋喃唑酮代谢物直接竞争化学发光检测方法的建立

目的:建立呋喃唑酮代谢物(AOZ)直接竞争化学发光检测方法,用于新疆水产品呋喃唑酮残留检测。方法:采用碳二亚胺法将AOZ与对醛基苯甲酸反应生成物CPAOZ与HRP偶联成酶标物CPAOZ-HRP,加之CPAOZ单克隆抗体,建立AOZ直接竞争化学发光检测方法(CLIA);运用高效液相色谱法(HPLC)和AOZ-CLIA检测新疆罗非鱼样品中AOZ的残留情况。结果:以1∶6000CPZOA单克隆抗体稀释度为最佳包被浓度,1∶64000CPAOZ-HRP稀释浓度为最佳检测浓度,建立AOZ-CLIA方法,其标准曲线呈线性,线性范围0.0527~8.5ng/mL,IC50为0.235ng/mL;该方法能特异性检测AOZ,与其他药物无交叉反应;样品最低检出限为0.0509μg/kg,样品添加回收率为87.2%~95.0%;新疆罗非鱼AOZ-CLIA检测结果与HPLC结果完全一致,均未检测出超标样本。结论:建立的AOZ-CLIA检测方法能满足新疆水产品呋喃唑酮残留检测。

Determination of furazolidone metabolite(AOZ)by direct competitive chemiluminescence immunoassay (CLIA)

CPAOZ is synthesized from AOZ and 4-Formylbenzoic acid by using carbon diimide method,and coupled with HRP to form HRP enzyme-marked conjugates(CPAOZ-HRP).Residue of AOZ in tilapias was detected by AOZ-CLIA.The optimum coating concentration and best detection concentration of CPAOZ-HRP were 1:6000 and 1:64000,respectively.The standard curve was linear in the range of0.0527-8.5 ng/mL,the 50%inhibitory concentration(IC50)was 0.235 ng/mL.AOZ could be specifically detected,was no cross-reaction with other furazolidone metabolite.The minimum detection limit was0.0509 ug/kg,the recovery rate of the spiked sample was 87.2%-95.0%.The determination results are completely consistent with that of HPLC method.