高效液相色谱串联质谱法测定人血浆中吲达帕胺浓度及生物等效性研究

Determination of Indapamide in human plasma by liquid chromatography tandem mass spectrum and study on its bioequivalence

目的:建立一种简单、有效且灵敏的液质联用法(LC-MS/MS)检测人血浆中吲达帕胺浓度。方法:血浆样品经叔丁基甲基醚(tert-ButylMethyl Ether,MTBE)萃取后,选安定作内标。色谱柱为Luna C18柱(150mm×2.0mm,5μm);流动相为10mmol/L甲酸铵(含0.1%甲酸):甲醇=(20∶80,V/V);流速为0.30mL/min;采用ESI+MRM方式监测;源电压:3.5kV;源温度:100℃;吲达帕胺和安定的离子选择通道分别为:m/z 366.2→132.1,285.2→154.1。结果:吲达帕胺线性范围为0.536～45.733ng/mL,最低检测浓度为0.536ng/mL,提取回收率在69%～81%,日内、日间RSD均<15%。由湖南协力药业有限公司生产生产的吲达帕胺缓释片(1.5mg/片)与施维雅(天津)制药有限公司生产的吲达帕胺缓释片(商品名:纳催离,1.5mg/片)经单次及多次给药试验研究,具有生物等效性。结论:本方法简单、灵敏,可准确检测人体血浆中吲达帕胺的浓度。

AIM： To establish a simple,effective and sensitive LC-MS/MS assay to determine Indapamide in human plasma.METHODS： Diazepam was used as an internal standard.Human plasma samples were extracted by MTBE（tert-Butyl Methyl Ether）.The separation was carried out on a Luna C18（150 mm×2.0 mm,5 μm） column.The mobile phase consisted of 10 mmol/L ammonium formate with 0.1% formic acid：methanol（20∶80,V/V） at a flow rate of 0.30 mL/min.Electrospray ionization（ESI） source was applied and operated in the positive ion mode.The MS detection parameters were as follows： the source voltage was 3.5 KV and source temperature was 100 ℃.Quantitation was performed using multiple reaction monitoring（MRM） of the m/z 366.2→132.1 for Indapamide and 285.2→154.1 for diazepam.RESULTS：The liner calibration curves were obtained in the range of 0.536-45.733 ng/mL for Indapamide.The lower limit of quantification was 0.536 ng/mL.The mean relative recovery was 69%~81% and the relative standard deviation of the intra-day and inter-day precision of variation were less than 15%.By single and multiple doses studies,we can demonstrate that Indapamide sustained release tablets（1.5 mg/tablet） produced by Hunan Xieli Pharmaceutical Co.,Ltd and Servier（Tianjin） Pharmaceutical Co.,Ltd are bioequivalent.CONCLUSION： This method which is simple and sensitive can be used to determinate the concentration of Indapamide in human plasma.