LC-MS/MS法同时测定人血浆中卢帕他定及其活性代谢产物地洛他定的浓度

Simultaneous determination of rupatadine and desloratadine concentration in human plasma by LC-MS / MS

目的建立同时测定人血浆中卢帕他定及其主要活性代谢产物地洛他定浓度的LC-MS/MS法。方法以艾司唑仑为内标,血浆样品经环己烷-乙醚(50∶50,V/V)提取后,以甲醇-乙酸铵溶液(5 mmol/L,p H=2.2)(70∶30,V/V)为流动相,采用Shimadzu ODS色谱柱(150 mm×2.0 mm,5μm)分离,电喷雾离子化-正离子选择性检测方式,检测离子为m/z:416.1→309.0(卢帕他定),311.2→295.0(地洛他定),295.0→267.1(内标艾司唑仑)。结果卢帕他定浓度在0.1 ng/ml^100.0 ng/ml、地洛他定浓度在0.1 ng/ml^50.0 ng/ml线性关系良好(r>0.99),最低检测限浓度均达0.05 ng/ml,高、中、低3种浓度的平均提取回收率均>70%,批内、批间精密度良好(RSD<15%)。结论该方法样品前处理简便、灵敏度高、精密度好、线性范围宽,为卢帕他定大批量生物样本的分析提供了新的选择。

Objective To establish a liquid chromatograph- Mass spectrum（ LC- MS / MS） method for simultaneous determination of rupatadine and desloratadine concentration in human plasma. Methods With estazolam as internal standard,rupatadine and desloratadine were extracted from plasma with cyclohexane- diethyl ether（ 50∶50,V / V）. An aliquot was chromatographically analyzed on the Shimadzu VP- ODS（ 150 mm × 2. 0 mm,5 μm） using the mobile phase comprised methanol- ammonium acetate（ 5 mmol / L,p H = 2. 2）（ 70 ∶30,V / V） by means of selective reaction monitoring（ SRM） mode mass spectrometry. The determination ion was respectively m / z： 416. 1→309. 0（ rupatadine）,311. 2 →295. 0（ desloratadine）,295. 0 →267. 1（ estazolam）. Results The linearity was good when the concentration of rupatadine was 0. 1 ng / ml ～ 100. 0 ng / ml and the concentration of desloratadine was 0. 1 ng / ml ～ 50. 0 ng / ml（ r〉 0. 99）. The lowest limit of detection concentration of the two compounds were 0. 05 ng / ml. The absolute extraction recovery was above 70%. The intra- batch precision and inter-batch precision in the method were good（ RSD〈 15%）. Conclusion This method was easy and convenient,high sensitive,accurate and good linear in the pre- treatment of samples,which provided a new choice for the analysis of large bio- samples,such as rupatadine.