可视化蛋白芯片法同时检测饲料中多种抗生素

Simultaneous determination of various antibiotics in feed by visual protein chip method

目的建立可视化蛋白芯片法同时检测饲料中的四环素、林可霉素、氟苯尼考等抗生素。方法采用戊二醛法合成四环素人工抗原,琥珀酸酐法合成林可霉素和氟苯尼考人工抗原,并用紫外分光光度法进行鉴定。通过优化最佳的抗原抗体浓度,计算3者相互之间的交叉反应率,建立标准曲线,分别考察饲料中四环素、林可霉素和氟苯尼考的加标回收率和重复性。结果方法的加标回收率均在70%~120%内,RSD均小于15%。用建立的方法应用于未知饲料样品的检测,结果与液相色谱-质谱联用(high performance liquid chromatography-mass spectrometry, HPLC-MS)的方法一致,相关系数均大于0.99。结论可视化蛋白芯片法具有检测时间短、操作简单、可进行大量样本同时检测,且一次可检测多个指标等优点,可满足饲料样本中多种抗生素检测的需求。

Objective To establish a method for simultaneous determination of tetracycline,lincomycin,fluorophenicol and other antibiotics in feed by visual protein chip method.Methods Tetracycline artificial antigen was synthesized by glutaraldehyde method,and lincomycin as well as florfenicol artificial antigen was prepared by succinic anhydride approach,and products were verified by ultraviolet spectrophotometer.By optimizing antigen-antibody concentration and calculating cross-reaction rate among them,the standard curve was established,and the recovery and repeatability of spiked tetracycline,lincomycin and florfenicol in the feed were investigated respectively.Results The spike recoveries were within 70%–120%,and RSDs were less than 15%.At the same time,the established method was applied to detect unknown feed samples.The results were consistent with HPLC-MS method,and correlation coefficients were all greater than 0.99.Conclusion The visual protein chip method has the advantages of short detection time,simple operation,simultaneous detection of a large number of samples,and multiple indicators at a time,which can meet the detection of multiple antibiotics in feed samples.