时间分辨荧光免疫层析定量检测牦牛肉中喹诺酮类药物

Quantitative Detection of Quinolone by Time-resolved Fluorescence Immunochromatography in Yak Meat

目的:本研究旨在建立一种简单、快速、高灵敏的牦牛肉中喹诺酮类药物(环丙沙星、恩诺沙星、诺氟沙星、氧氟沙星、达氟沙星等)残留的荧光定量免疫层析检测方法。方法:将环丙沙星鼠单克隆抗体G10785与羧基化铕微球进行偶联标记,环丙沙星抗原和羊抗鼠二抗分别包被于硝酸纤维素膜(NC)上作为检测线(T)和控制线(C);根据T线荧光信号值与C线荧光信号值的比值(T/C)和样本中喹诺酮类药物浓度建立定量标准曲线;对所建立方法的灵敏度、准确度、精密度等进行评价,并将其与LC-MS/MS方法进行对比。结果:本研究所建立方法的定量限<0.96μg/kg,批内回收率为82.94%~111.27%,变异系数为4.13%~9.90%,批间回收率为88.86%~107.05%,变异系数为3.86%~9.39%。与环丙沙星、恩诺沙星、诺氟沙星、氧氟沙星、达氟沙星的交叉反应率分别为100%、77.36%、107.66%、93.55%和50.92%。结论:本研究所建立的时间分辨荧光免疫层析检测方法准确度、精密度、灵敏度较高,性能稳定,具有良好的应用前景。

Objective:The purpose of this study was to establish a simple,rapid and highly sensitive immunochromatographic method for the determination of quinolones(ciprofloxacin,enrofloxacin,norfloxacin,levofloxacin,and so on)in yak meat.Methods:Ciprofloxacin mouse monoclonal antibody G10785 was coupled with carboxylated europium microspheres,and ciprofloxacin antigen and goat anti-mouse antibody were coated on cellulose nitrate membrane(NC)as detection line(T)and control line(C),respectively.Quantitative standard curves were established according to the value of T/C and quinolone concentration in samples.The sensitivity,accuracy,and precision of the method were evaluated,and the yak meat samples were analyzed by using both the developed method and LC-MS/MS.Results:The limit of quantitation of the established method was lower than 0.96μg/kg,the in-batch recovery rate ranged from 82.94%to 111.27%,and the coefficient of variation ranged from 4.13%to 9.90%,the inter-assay recovery rate ranged from 88.86%~107.05%,and the coefficient of variation ranged from 3.86%~9.39%.The cross-reactivity rates of the established method with ciprofloxacin,enrofloxacin,norfloxacin,levofloxacin,and danofloxacin were 100%,77.36%,107.66%,93.55%,and 50.92%,respectively.Conclusion:The time-resolved fluorescence immunochromatographic method established in this study had high accuracy,precision,sensitivity and stable performance,and had a good application prospect.