高效液相色谱紫外电化学串联检测生物样品中克仑特罗残留

Determination of clenbuterol in biological samples by high performance liquid chromatography with in seies ultraviolet and electrochemical detection

采用高效液相色谱紫外电化学串联法测定了动物性组织和体液中克仑特罗残留的含量。样品经初提 ,液液萃取 ,弱阳离子交换柱分离富集浓缩后经高效液相色谱分离 ,紫外电化学检测器串联检测。液相色谱的流动相为甲醇 :0 0 1mol LKCl溶液 (4 5 :5 5 )。色谱柱为HypersilBDSC18柱。紫外检测波长 2 4 4nm ,电化学采取脉冲方式 ,峰电压 1 0V。克仑特罗浓度与紫外及电化学响应信号均呈良好的线性关系。不同加标水平的回收率在 4 2 2 %～ 96 1%之间。样品的定量限为 0 5 μg kg。对猪肉、猪肝、猪肾、鸡肉和尿液、血液等生物样品进行了测定 。

A method for the determination of clenbuterol in animal tissues and fluid is described. After a three-step pretreatment, involving sample extract, liquid-liquid partition and purification on a LC-WCX column, the separation of clenbuterol from interfering compounds present in biological samples was performed on a Hypersil BDS C18 column by high performance chromatography with in series ultraviolet and electrochemical deterction. The mobile phase was a mixture of methanol and 0.01mol/L potassium chloride(45∶55). The UV-absorption 244nm and the electrochemical detection pulse mode +1.0V were applied in series. The results showed that concentration of clenbuterol and peak areas achieved a nice linear relation. The limit of detection of clenbuterol in animal foods is 0.5μg/kg. The mean recovery of clenbuterol spiked at 2.5,2.0,1.5,0.5μg/kg levels with chicken and liver samples were 42.2%-96.1%, RSD were from 12.4 to 19.7%(n=7). With the described method, animal food samples and biological fluid(e.g.urine and blood) were analyzed with satisfaction.