液相色谱串接质谱联用法确证肉食品中低浓度克仑特罗

Determination of Low-concentration Clenbuterol in Edible Meat Using High Performance Liquid Chromatography-tandem Mass Spectrometry

目的:采用液相色谱串接质谱联用法对肉食品中残留的低浓度克仑特罗进行确证分析。方法:样品匀浆处理后,酸化去除蛋白,经液液萃取和MCX 3cc固相萃取柱两步纯化,以流动相为10 mmol/L的p H 3.5甲酸铵和乙腈为流动相,梯度洗脱,采用Eclipse C18(1.8μm,4.6×100 mm)色谱柱分离,电喷雾离子源,正离子多反应监测模式扫描分析检测。结果:D9-克仑特罗为内标,克仑特罗的线性范围为0.01~0.2μg/kg,相关系数(R2)大于0.99,检出限为0.005μg/kg,3个不同水平的加标回收率为78.8%~114.8%,相对标准偏差不大于10%。结论:该方法具有操作简单、灵敏度高、重现性好等特点,可以完成肉食品样品中痕量克仑特罗的确证分析。

Objective To determine low-concentration clenbuterol in edible meat based on the solid- phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）. Method The homogenated sample was acidized to remove proteins,and purified using the liquid-liquid extraction and MCX Oasis solid prepared extraction column,then further treated with gradient elution with the mobile phase of ammonium formate （10 mmol/L and pH 3.5） and acetonitrile. The clenbuterol was completely separated on Eclipse C18 （1.8 μm,4.6×100 mm） column and detected in multiple reaction monitoring（MRM）mode. Results A good linearity was achieved for clenbuterol in the arrange of 0.01-0.2 μg/kg based on the internal standard calibration of D9- clenbuterol, with the linearity correlation coefficient greater than 0.99 and the detection limit of 0.005 μg/kg. The relative recovery of target compounds spiked in blank sample at three levels ranging from 78.8 to 114.8% ,with the relative standard deviations less than 10%. Conclusion The method in this research is simple,rapid,reliable and suitable to confirm low-concentration clenbuterol in edible meat.