HPLC-MS/MS法同时测定牛肉中80种兽药及其代谢物

Determination of 80 Veterinary Drug Residues and Their Metabolites in Beef by HPLC-MS/MS

为全面排查宁夏地区牛肉兽药残留风险,从农业农村部规定的禁停用药物、GB 31650—2019《食品安全国家标准食品中兽药最大残留限量》中规定有最大残留限量的药物以及宁夏养殖环节大量使用药物三方面筛选确定12类药物,利用简单、快捷的前处理方法和超高效液相色谱-串联质谱多反应监测分析技术,建立了一次进样可同时分析检测12类80种兽药及其代谢物残留的快速检测方法。样品采用Oasis PRiME HLB固相萃取柱净化,经Waters HSS T3色谱柱分离,多反应监测模式监测,基质匹配外标法定量。结果表明:80种兽药及其代谢物在基质匹配标准溶液1.0~50.0μg/L的质量浓度范围内呈现良好的线性关系,相关系数均大于0.990,方法定量限在2.0~5.0μg/kg,方法在2.0~100.0μg/kg的平均回收率为60%~103%,相对标准偏差(n=5)均小于20%;应用该检测方法对40份市售牛肉进行检测,其中4份牛肉检出兽药残留,分别为土霉素(54.5μg/kg)、磺胺间甲氧嘧啶(23.3μg/kg和35.4μg/kg)及恩诺沙星(118μg/kg),再分别使用GB/T 21317—2007、GB/T 20759—2006、GB/T 21312—2007方法对阳性样品进行复检,结果一致。经验证表明:该检测方法定性定量结果准确可靠、灵敏度高、重现性好,可满足对牛肉中兽药残留检测分析的要求。

In order to comprehensively investigate the risk of veterinary drug residue in beef in Ningxia,in this study,12 types of drugs were screened and identified from three aspects:banned and stopped drugs prescribed by the Ministry of Agriculture and Rural Affairs,the drugs with the maximum residue limits specified in the GB 31650—2019“National Food Safety Standards Maximum Residue Limits of Veterinary Drugs in Food”,and drugs used in large quantities in Ningxia's breeding process.Using simple and fast pre-processing methods and high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)multi-reaction monitoring and analysis technology,a rapid detection method was established which could simultaneously analyze and detect 12 types of 80 veterinary drugs and their metabolites with one injection.The sample was purified by Oasis PRiME HLB solid phase extraction column,separated by Waters HSS T3 chromatographic column,monitored by multi-reaction monitoring mode,and quantified by matrix matching external standard method.The results showed that 80 veterinary drugs and their metabolites showed a good linear relationship in the concentration range of matrix matching standard solution 1.0-50.0μg/L,with the correlation coefficients greater than 0.990.The quantitative limits of the method was 2.0-5.0μg/kg,and the average recoveries in the range of 2.0-100.0μg/kg were 60%-103%,and the relative standard deviation was less than 20%(n=5).This method was used to detect 40 pieces of commercially available beef,of which 4 pieces of beef were detected with veterinary drug residues,including oxytetracycline(54.5μg/kg),sulfamethoxine(23.3μg/kg and 35.4μg/kg),and enrofloxacin(118μg/kg).The positive samples were re-examined using GB/T 21317—2007,GB/T 20759—2006,and GB/T 21312—2007 methods,and the results were consistent.The verification showed that the qualitative and quantitative results of the detection method were accurate,reliable,sensitive,and reproducible,and could satisfy the requirements for the detection and analysis of veterinary drug residues in beef.