LC-MS法测定人血浆中比索洛尔的浓度

LC-MS determination of bisoprolol in human plasma

目的:建立高效液相色谱-质谱联用法测定人血浆中比索洛尔浓度的方法,并研究富马酸比索洛尔片剂在健康人体的药代动力学。方法:血浆样品以阿奇霉素作为内标,经2mol·L-1氢氧化钠溶液0.1mL碱化后乙酸乙酯萃取进行LC-MS分析。色谱条件为KromasilC18柱(150mm×4.6mm,5μm),流动相为甲醇-0.2%甲酸(55∶45,v/v),流速0.8mL.min-1;质谱条件为电喷雾离子源(ESI源),正离子方式检测:扫描方式为选择离子监测(SIM),用于定量分析的离子分别为m/z326.25(比索洛尔)和m/z749.80(阿奇霉素)。结果:比索洛尔浓度在0.2～150.0ng·mL-1范围内线性关系良好,方法的定量下限(LLOQ)为0.2ng·mL-1,日内、日间精密度(RSD)均小于13.2%,提取回收率在95.2%～98.2%之间。结论:该方法简便、准确,灵敏度高,专属性强,适用于人血浆中比索洛尔浓度的测定。

Objective：To develop an LC-MS method for the determination of bisoprolol concentration in human plasma and study the pharmacokinetics of bisoprolol in healthy volunteers after a single oral dose of 5 mg bisoprolol fumarate tablet preparation.Method：After adding azithromycin as the internal standard and 0.1 mL 2 mol·L-1NaOH as the basification reagent,bisoprolol was extracted from plasma by ethyl acetate and the separation was carried on a Kromasil C18 column （150 mm×4.6 mm,5 μm）with a mobile phase of methanol-0.2% formic acid（55：45） at a flow rate of 0.8 mL·min-1.Bisoprolol was determined by LC-ESI/MS in selected ion monitoring（SIM） mode,the ion combination of m/z 326.25 and m/z 749.80 was used to qualify bisoprolol and the internal standard,respectively.Result：The calibration curve was in good linearity over the range of 0.2-150.0 ng·mL-1.The lower limit of quantification（LLOQ） for bisoprolol in plasma was 0.2 ng·mL-1.The intra-day and inter-day precision （RSD） were lower than 13.2%.The recovery for bisoprolol was between 95.2%-98.2%.Conclusion：The method is shown to be simple,accurate,sensitive and specific for the determination of bisoprolol in human plasma.