高通量微生物显色法快速检测动物源性食品中抗生素残留  

Rapid and High-Throughput Detection of Antibiotic Residues in Animal-Derived Food Samples by Microbial Chromotest

作　　者：范维[1] 高晓月[1] 李贺楠[1] 李莹莹[1] 郭文萍[1] 陈淑敏[1] FAN Wei;GAO Xiaoyue;LI Henan;LI Yingying;GUO Wenping;CHEN Shumin(China Meat Research Center, Beijing Academy of Food Science, Beijing 100068, China)

机构地区：[1]中国肉类食品综合研究中心,北京食品科学研究院,北京100068

出　　处：《食品科学》2017年第16期239-244,共6页Food Science

基　　金：“十二五”国家科技支撑计划项目(2014BAD04B05)

摘　　要：建立一种采用高通量微生物显色法对动物源性食品中抗生素残留进行筛检,再结合高效液相色谱-串联质谱(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)法对阳性样品进行复测的分析检验方法。本方法利用嗜热脂肪芽孢杆菌(Bacillus sterothermophilus)作为指示菌,制备96微孔板,可同时对不同类别多种抗生素进行联合检测。结果表明:该微生物显色法操作简便、快捷,成本低,结果准确且易判断,选取pH 7.4磷酸盐缓冲液进行提取,以反应液150μL、菌液50μL(初始OD_(600 nm)为0.4左右)、样品提取液100μL作为检测体系,检测效果最佳,对动物源性食品中氨基糖苷类检出限为60μg/kg,β-内酰胺类检出限为20~40μg/kg,大环内酯类检出限为60~80μg/kg,四环素类检出限为40~60μg/kg,符合国内外对抗生素残留限量的要求。对60份动物源性样品进行检测,其结果与HPLC-MS/MS复测结果一致,说明该微生物法稳定性良好,可用于动物源性食品中抗生素残留的初筛检测。A new rapid and high-throughput microbial chromogenic assay was developed to analyze antibiotic residues in animal-derived food samples.The positive samples were further confirmed by high performance liquid chromatographytandem mass spectrometry(HPLC-MS/MS)method.Bacillus sterothermophilus was used as an indicator strain to produce a96-well microplate for detecting a variety of antibiotics simultaneously.The results showed that the microbial assay was simple and cheap,and gave easy-to-interpret results.After extraction with phosphate buffer at pH7.4,muscle samples were detected using a chromogenic assay system consisting of the reactant solution(150μL),bacterial suspension(50μL)with an initial optimal density(OD600nm)of around0.4,and the sample extract(100μL).The limits of detection(LODs)of aminoglycosides,beta-lactam,macrolide,and tetracyclines antibiotics were60,20-40,60-80,and40-60μg/kg,respectively,which reached the requirements for the determination of antibiotic residue levels in the country and abroad.Consistent results were obtained in examining antibiotic residue in60animal-derived food samples using the microbial chromogenic assay and HPLC-MS/MS,indicating that the proposed assay could be used to detect antibiotic residues in animal-derived food samples with reliable results.

关 键 词：高通量微生物显色法 抗生素残留 高效液相色谱-串联质谱 动物源性食品 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]