LC-MS/MS法测定人血浆中氨溴索浓度及其生物等效性研究

Determination of Ambroxol in Human Plasma by LC-MS/MS:Application in a Bioequivalence Study

目的:建立LC-MS/MS法测定人血浆中氨溴索浓度,并进行生物等效性研究。方法:血浆样品用乙腈沉淀蛋白,内标为卡马西平,采用电喷雾化离子源,监测的氨溴索和内标的离子对分别为m/z379.0/263.9和237.0/194.1,色谱柱为UltimateXB-CN(2.1 mm×150 mm,5μm),流动相为乙腈:0.2%甲酸=40:60,流速为0.3 ml·min^(-1),进样量为10μl。结果:氨溴索标准曲线线性范围为0.53～187.50 ng·ml^(-1),最低定量限为0.53 ng·ml^(-1),低、中、高三种浓度的批内和批间精密度RSD(%)均小于15%,准确度分别为101.41%、105.40%和106.04%,提取回收率分别为82.51%、93.57%和89.19%,基质效应分别为90.48%、92.36%和100.81%,稳定性试验表明样品在本试验条件下稳定。结论:该方法灵敏度高、专属性强、准确、简便,适用于氨溴索的生物等效性研究。

Objective： To develop an ultra performance liquid chromatography-tandem mass spectrometry method （LC-MS/MS） for the determination of ambroxol in human plasma and its bioequivalenee study. Method ： Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization. The ion pairs for ambroxol and internal standard were monitored at m/z 379. 0/263.9 and 237.0/194. 1 in positive ion mode, respectively. Carbamazepine was used as the internal standard, and aeetonitrile was used to precipitate plasma proteins. The column was Ultimate XB-CN （2. 1 mm ×150 mm,5 μm）. The mobile phase consisted of acetonitrile and 0. 2% formic acid （40： 60） at a flow rate of 0. 3 ml.min-1. Result： Linearity was within the range of 0. 53-187.50 ng · ml^-1. The lowest quantitation limit was identifiable and reproducible at 0. 53 ng·ml^-1. The Intra- and inter-day RSD were both less than 15%. The accuracy was 101.41% , 105.40% and 106. 04% ,respectively. The extraction recoveries were 82. 51% ,93.57% and 89. 19% , and the matrix effects were 90. 48 % , 92. 36% and 100. 81% , respectively. The stability tests showed the promising stability of the samples during the whole study. Conclusion： A rapid, sensitive and simple LC-MS/MS method for determination of ambroxol in human plasma is developed, which is successfully applied in its bioequivalence study.