HPLC-MS/MS法测定猪肉中克伦特罗和莱克多巴胺

Determination of Clenbuterol and Ractopamine in Pork by High Performance Liquid Chromatography-Tandem Mass Spectrometry

研发聚合物整体柱微萃取(Polymer monolith microextraction,PMME)柱,建立猪肉中克伦特罗和莱克多巴胺的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法。样品用乙酸铵缓冲液提取,通过聚(甲基丙烯酸-乙二醇二甲基丙烯酸酯(p(MAA-co-EGDMA))柱进行富集净化,用甲醇洗脱,采用HPLC-MS/MS进行测定,梯度洗脱,流动相为甲醇和含0.1%的甲酸水溶液;质谱采集模式为电喷雾正离子多反应监测模式。以克伦特罗-D9为内标,克伦特罗的的线性范围为0.015μg/L^1.00μg/L,相关系数(R2)大于0.99,定量限为0.05μg/kg,3个不同水平的加标平均回收率为88.9%~105.0%,相对标准偏差小于10%。以莱克多巴胺-D3为内标,莱克多巴胺的的线性范围为0.15μg/L^10.0μg/L,相关系数(R2)大于0.99,定量限为0.50μg/kg,3个不同水平的加标平均回收率为89.9%~105.0%,相对标准偏差小于10%。该方法具有操作简单、灵敏度高、重现性好,试剂用量少等特点,符合食品样品中痕量污染物的检测要求。

A method was established for the determination of clenbuterol and ractopamine in pork based on Polymer monolith microextraction coupled with high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS）.The samples was extracted by ammonium acetate buffer and further purified by an PMME,which was eluted by methanol.The target compounds were assayed by HPLC-MS/MS with gradient elution using methanol and 0.1 % of formic acid aqueous solution as mobile phases.The mass spectrometric acquisitions were carried out by means of multiple reaction monitoring（MRM） in electrospray positive ionization mode. The clenbuterol linearities（R20.99）were achieved over the range of 0.015 μg/L-1.00 μg/L based on the internal standard calibration of clenbuterol-D9,the quantification limits of the method were 0.05 μg/kg.The mean recoveries of the clenbuterol（spiked at three concentration levels）ranged from 88.9 % to 105.0 %,with the relative standard deviations（RSDs） no more than 10 %. The ractopamine linearities（R2 0.99） were achieved over the range of 0.15 μg/L-10.0 μg/L based on the internal standard calibration of ractopamine-D3,the quantification limits of the method were 0.50 μg/kg.The mean recoveries of the clenbuterol（spiked at three concentration levels）ranged from 89.9 % to 105.0 %,with the relative standard deviations（RSDs）no more than 10 %. This proposed method is simple,sensitive,reproducible,and complied with the regulations for the determination of trace contaminants residues in pork matrices.