超高效液相色谱-串联质谱法测定猪肝中4种四环素类药物残留

Determination of 4 kinds of tetracyclines residues in swine liver by ultra performance liquid chromatography-tandem mass spectrometry

目的建立超高效液相色谱-串联质谱法同时测定猪肝中4种四环素类药物残留量的分析方法。方法样品经弱酸性的Na2EDTA-Mcllvaine缓冲溶液提取,硫酸溶液与钨酸钠溶液沉淀蛋白,HLB固相萃取柱净化,WatersACQUITYHSST3色谱柱分离,0.1%甲酸水溶液与乙腈作为流动相梯度洗脱,多反应监测正离子模式测定,内标法定量。结果 4种四环素类药物在5~100 ng/mL范围内线性关系良好,相关系数r^(2)均大于0.990,定量限为10μg/kg,在10~50μg/kg添加水平上平均回收率在95.4%~111.2%范围内,批内与批间相对标准偏差均小于10.1%。结论该方法简便快速、结果准确、回收率高,有效地克服了基质干扰与操作损耗对定量结果的影响,适合大批量猪肝样品的快速定量分析。

Objective To establish an analytical method for the simultaneous determination of 4 kinds of tetracyclines residues in swine liver by ultra performance liquid chromatography-tandem mass spectrometry.Methods The samples were extracted with weakly acidic Na 2 EDTA-Mcllvaine buffer solution,followed by protein precipitation with sulfuric acid solution and sodium tungstate solution,purified with HLB solid-phase extraction column,and separated with Waters ACQUITY HSS T3 chromatographic column.Gradient elution was performed with 0.1%formic acid aqueous solution and acetonitrile as the mobile phase,multi-reaction monitoring positive ion mode was used for determination,and internal standard method was used for quantification.Results The linear relationships of the 4 kinds of tetracyclines were good within the range of 5‒100 ng/mL,with correlation coefficients r^(2)>0.990,and the limits of quantitation were 10μg/kg.The average recoveries were within the range of 95.4%to 111.2%at the level of 10‒50μg/kg,and the relative standard deviations were less than 10.1%within and between batches.Conclusion This method is simple,rapid,accurate and has high recovery rate,which effectively overcomes the influence of matrix interference and operation loss on quantitative results,and is suitable for rapid quantitative analysis of large quantities of swine liver samples.