UPLC-MS/MS法测定动物源性食品中氯苯胍及其代谢物残留  

Determination of Robenidine and Its Metabolites in Animal Derived Food by UPLC-MS/MS

作　　者：郑幸果 郭灵安[1] 赵珊[1] 李曦[1] 雷欣宇[1] 仲伶俐[1] ZHENG Xingguo;GUO Lingan;ZHAO Shan;LI Xi;LEI Xinyu;ZHONG Lingli(Analysis and Testing Center of Sichuan Academy of Agricultural Sciences,Chengdu 610066,China)

机构地区：[1]四川省农业科学院分析测试中心,成都610066

出　　处：《中国畜牧兽医》2020年第6期1961-1970,共10页China Animal Husbandry & Veterinary Medicine

基　　金：四川省应用基础项目(2019YJ0611)。

摘　　要：为快速有效地检测动物源性食品中氯苯胍及其代谢物残留,研究建立一种在鸡蛋、鸡肉、牛肉、鱼肉和猪肉5种动物源性食品中,同时检测氯苯胍及其代谢物(对氯苯甲酸、对氯苯甲酰氨基乙酸)残留的超高效液相色谱-串联质谱(UPLC-MS/MS)法。样品经过2%(V/V)甲酸乙腈溶液提取,无水硫酸钠去除水分,氮吹浓缩后甲醇复溶,正己烷除脂,高速冷冻离心,得到净化后的样品进行上机测定。选用Waters ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),将甲醇—0.1%甲酸水溶液作为流动相进行梯度洗脱。通过多反应检测(MRM),在正/负离子模式下,采用基质匹配外标法,同时对3种化合物进行定性和定量分析。结果显示,氯苯胍、对氯苯甲酸和对氯苯甲酰氨基乙酸在各自浓度范围内线性关系良好,相关系数R2>0.999。氯苯胍的检出限(LOD)和定量限(LOQ)分别为0.5和1.0μg/kg,对氯苯甲酸的LOD和LOQ分别为2.5和5.0μg/kg,对氯苯甲酰氨基乙酸的LOD和LOQ分别为1.0和2.5μg/kg。不同基质中,3种化合物在4个添加水平(氯苯胍:1.0、25、50、100μg/kg;对氯苯甲酸:5.0、25、50、100μg/kg;对氯苯甲酰氨基乙酸:2.5、25、50、100μg/kg)的平均回收率为76.0%~95.9%,相对标准偏差(RSDs,n=6)为2.6%~10.6%。基质效应|ME|为0.2%~26.2%,其中氯苯胍在鸡蛋中,对氯苯甲酰氨基乙酸在鸡肉、牛肉和鱼肉中存在较强的基质效应(|ME|>20%),空白猪肉可作为代表基质用于3种化合物的定量分析。本方法前处理简单,灵敏度较高,重现性好,可用于动物源食品中氯苯胍及其代谢物残留的测定。In order to detect robenidine and its metabolites residues in animal derived food rapidly and effectively,an ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed for the simultaneous determination of robenidine and its metabolites(4-chlorobenzoic acid,4-chlorobenzoylamino-acetic acid)residues in five kinds of animal derived food,including eggs,chicken,beef,fish and pork.Samples were extracted with 2%(V/V)formic acid-acetonitrile solution,removed water with anhydrous sodium sulfate,fixed volume with methanol,then excepted fat with n-hexane.The purified samples were analyzed by UPLC-MS/MS after freezed and high speed centrifuged.UPLC was achieved using a Waters ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7μm);the mobile phase consisted of methanol and 0.1%formic acid-water solution.The qualitative analysis was performed with MS/MS under multiple reaction monitoring(MRM)mode with positive and negative electrospray ionization;the matrix matched external standard method was used for quantitation.The results showed that there were good linearities of the three compounds in their respective concentration range(R2>0.999).The limits of detection(LODs)and the limits of quantity(LOQs)for robenidine were 0.5 and 1.0μg/kg,for 4-chlorobenzoic acid were 2.5 and 5.0μg/kg,for 4-chlorobenzoylamino-acetic acid were 1.0 and 2.5μg/kg,respectively.While blank samples spiked four levels(robenidine:1.0,25,50,100μg/kg;4-chlorobenzoic acid:5.0,25,50,100μg/kg;4-chlorobenzoylamino-acetic acid:2.5,25,50,100μg/kg),the recoveries and RSDs(n=6)of the three compounds were 76.0%-95.9%and 2.6%-10.6%,respectively.The matrix effects(|ME|)of the developed method were 0.2%-26.2%.There was a strong matrix effect(|ME|>20%)of robenidine in eggs and 4-Chlorobenzoylamino-acetic acid in chicken,beef and fish;blank pork could be used as representative matrix for quantitative analysis of three compounds.Therefore,the method was simple in pretreatment,high in sensitivity and good in reproducibility,and could

关 键 词：超高效液相色谱-串联质谱(UPLC-MS/MS) 动物源性食品 氯苯胍 代谢物 

分 类 号：S859.84[农业科学—临床兽医学]