LC-MS/MS测定人血浆中阿米舒必利的浓度

Determination of Amisulpride in Plasma by LC–MS/MS

目的建立测定人血浆中阿米舒必利浓度的高效液相色谱串联质谱电喷雾检测法(LC-MS/MS)。方法以WelchMaterials XB-C18(2.1mm×150mm,3μm)为色谱柱,流动相为甲醇-水(95∶5,含5mmol·L-1甲酸铵),流速为0.3mL·min-1,柱温:40℃,以乙酸乙酯-二氯甲烷(4∶1)为提取溶剂。样品经电喷雾离子源正离子化后,通过三重四级杆串联质谱仪,采用选择反应监测(SRM)对阿米舒必利(m/z370.3→242.1)和内标舒必利(m/z324.2→112.1)进行测定。结果阿米舒必利高(400μg·L-1)、中(250μg·L-1)、低(1.25μg·L-1)3个浓度的平均方法回收率分别为104.44%、104.74%和95.65%,日内(n=5)、日间(n=3)RSD均小于15%;分析方法的最低定量限为0.52μg·L-1。线性范围为:0.5～500μg·L-1。结论该方法灵敏、准确、简单、快速,可用于阿米舒必利临床血浓监测和药动学研究。

OBJECTIVE To establish an LC-MS/MS method for the determination of amisulpride in human plasma. METHODS The sample extraction was performed by ethyl acetate-dichloromethane（4 ： 1）. Separation was achieved on a Welch Materials XB-C18（2.1 mm×150 mm, 3 μm） column with the mobile phase consisted of methanol-water with 5 mmol·L^-1 ammonium formate （95：5）. An Agilent 6410 triple quad mass spectrometer system equipped with an electrospray ionization ion-trap source was used as the detector and operated in the positive ion mode. Selected reaction monitoring（SRM） was used as the precursor to product ion combinations of m/z 370.3→242.1 and m/z 324.2→ 112.1 to detect amisulpride and the internal standard, respectively. The method was used to evaluate clinical application for 10 healthy volunteer after oral dose of 200 mg amisulpride tablet. RESULTS The average recoveries for amisulpride were 104.44%, 104.74% and 95.65%, respectively. The method was accurate with intra-day（n=5） and inter-day （n=3） RSD within 15%. The calibration curves for amisulpride had good linearity, over the concentration range of 0.5-500 μg·L^-1. The limits of quantitation for amisulpride was 0.52 μg·L^-1. CONCLUSION The method provides a sensitive, accurate, precise and reliable analytical procedure for clinical monitoring of amisulpride plasma and its phamacokinetic studies.