免疫亲和柱净化-UPLC-MS/MS测定鱼虾肉中的3种氨基糖苷类抗生素  

Determination of Three Aminoglycoside Antibiotics in Fish and Shrimp by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry with Immunoaffinity Column Clean-up

作　　者：王强[1] 王旭峰[1] 赵东豪[1] 黎智广[1] 蔡楠[1] 关婉琪 黄珂[1] 李刘冬[1] WANG Qiang;WANG Xufeng;ZHAO Donghao;LI Zhiguang;CAI Nan;GUAN Wanqi;HUANG Ke;LI Liudong(Key Laboratory of Aquatic Product Processing,Ministry of Agriculture,Laboratory of Quality and Safety Risky Assessment for Aquatic Product on Storage and Preservation（Guangzhou）,Ministry of Agriculture,Fishery Environment and Aquatic Products Quality Inspection and Testing Center（Guangzhou）,Ministry of Agriculture,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China)

机构地区：[1]中国水产科学研究院南海水产研究所农业部水产品加工重点实验室农业部水产品贮藏保鲜质量安全风险评估实验室(广州)农业部渔业环境及水产品质量监督检验测试中心(广州),广东广州510300

出　　处：《食品科学》2018年第20期326-331,共6页Food Science

基　　金：国家农产品质量安全风险评估重大专项(GJFP201501003;GJFP201700901);广东省水产品质量安全专项(GDSCZA2015008);中国水产科学研究院南海水产研究所基本科研业务费专项(2015TS17)

摘　　要：将庆大霉素(gentamicin,GEN)、卡那霉素(kanamycin,KAN)和安普霉素(apramycin,APR)的抗体同时偶联到溴化氰活化的琼脂糖凝胶上,制备成复合型免疫亲和柱(immunoaffinity column,IAC)。建立鱼虾肉中3种氨基糖苷类抗生素的免疫亲和柱净化-超高效液相色谱-串联质谱分析方法。采用BEH Amide(100 mm×2.1 mm,1.7μm)色谱柱分离,以0.05%甲酸溶液和乙腈为流动相梯度洗脱,电喷雾正离子多反应模式监测。结果表明,所制备的IAC对GEN、KAN和APR的柱容量分别为1 127、1 368、925 ng/mL,洗脱液选用0.1%甲酸-甲醇溶液。GEN的线性范围为80.0～500μg/L,KAN和APR的线性范围为20～500μg/L,相关系数均大于0.995。鱼虾肉基质中3种物质的平均回收率为71.7%～96.8%,相对标准偏差为4.2%～10.9%(n=6),检出限和定量限分别为10～40μg/kg和20～80μg/kg。为水产品中氨基糖苷类药物残留监控提供一种有效的技术手段。The polyclonal antibodies against gentamicin, kanamycin and apramycin were simultaneously coupled to CNBr-activated Sepharose 4B to make a composite immunoaffinity column （IAC）. An ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method was developed for the simultaneous determination of the three aminoglycosides （AGs） in fish and shrimp with immunoaffinity column clean-up. The analytes were separated on a BEH Amide column（100 mm × 2.1 mm, 1.7 μm）utilizing gradient elution with a mixture of acetonitrile and 0.05% formic acid solution as the mobile phase. The analysis was carried out in the multiple reaction monitoring （MRM） mode using positive ion electrospray ionization （ESI）. The column capacities of gentamicin, kanamycin and apramycin were 1 127, 1 368, and 925 ng/mL of gel, respectively. The optimum elution solvent was methanol containing 0.1% formic acid. The linear range of the method was 80.0–500 μg/L for gentamicin and 20–500 μg/L for kanamycin and apramycin, and the corresponding coefficients of correlation were all above 0.995. Recoveries from spiked fish and shrimp were in the range of 71.7%–96.8%, with relative standard deviations （RSDs） ranging from 4.2% to 10.9%. The limits of detection and the limits of quantitation were in the range of 10–40 and 20–80 μg/kg, respectively. The developed method would be a useful tool for monitoring aminoglycoside residues in aquatic products.

关 键 词：免疫亲和柱 超高效液相色谱-串联质谱 残留检测 氨基糖苷类抗生素 

分 类 号：O657.72[理学—分析化学]