超高效液相色谱-串联质谱法检测猪血清中泰乐菌素残留的建立

建立一种超高效液相色谱-串联质谱(UPLC-MS/MS)法测定猪血清中泰乐菌素的残留。血清样品在弱酸性条件下,经乙腈溶液提取,Sep-Vac C18固相萃取柱净化,用ACQUITY UPLC BEH-C18色谱柱(50 mm×2.1mm,1.7μm)进行分离,以乙腈-0.2%乙酸作流动相梯度洗脱,经电喷雾正离子模式,多反应监测(MRM)方式质谱分析,外标法定量。泰乐菌素含量在0.5~50 ng.mL-1范围内与峰面积μ呈良好线性,相关系数R2>0.9994,本方法的检测限为0.2 ng·mL-(1S/N>10)。样品添加浓度在5~50 ng·mL-1时,其回收率为73%~84%,日间与日内RSD均小于13.5%(n=5)。建立了一种快速、简单、廉价、高效和严格的方法来检测猪血清中的泰乐菌素残留情况,并为药物代谢动力学研究提供了可靠的检测方法。

Develop a method of ultra performance liquid chromatographytandem mass spectrometry to determinate Tylosin in swine serum

To establish a method of Ultra Performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) for determination of Tylosin in swine serum.The serum samples were deposited by acetonitrile directly in base,conditioning by Sep-Vac C18 Solid phase extraction(SPE) column.The analytes were separated by using ACQUITY UPLC BEH-C18(50 mm × 2.1 mm,1.7 μm) column,gradiented elution with the mobile phase consisted of acetonitrile-0.2% acetic acid.Determined by multiple reaction monitoring(MRM) mode with positive electrospray ionization(ESI +),quantitated with external standard method.Results showed that the calibration curve was linear in the range of 0.5～50 ng.mL-1,with coefficient of determination R2<0.9994.The minimal detectable serum drug concentration was 0.2 ng.mL-1(S/N>10).The average recovery was between 73% and 84%,at the range of 5～50 ngomL-1.Intra-day and inter-day RSDs were less than 13.5%(n=5).This study concerned the development of the quick,easy,cheap,effective and rugged(QuEChER) method for TYL,and provided a reliable detection method for Pharmacokinetic study.