LC-MS-MS法测定人血浆中非布司他浓度

Determination of febuxostate in human plasma by LC-MS-MS

目的建立液质联用色谱法（LC-MS-MS）测定人血浆中非布司他（febuxostate）浓度。方法空白血浆加非布司他,用乙腈作为沉淀剂,取上清液用于LC-MS-MS分析。分析柱为Thermo Biobasic-8柱（5μm,50 mm×2.1 mm）,流动相为乙腈-10 mmol/L乙酸铵（含0.05%甲酸）（70∶30）,流速为0.2 ml/min;质谱条件：电喷雾离子化电离源ESI负离子检测,喷雾电压（SP）3 500 kV,鞘气（SGP）流速10 Arb,辅助气（AGP）流速45 Arb,毛细管温度（TEM）270℃;非布司他和内标苯扎贝特的碰撞能量分别为10 eV、18 eV;选择反应监测（SRM）分别测定非布司他和内标苯扎贝特负离子m/z 315→271和m/z 360→274。结果非布司他在10-8 000μg/L检测浓度范围内呈良好线性关系（r〉0.99）,最低定量限（LLOQ）为10μg/L,绝对回收率在85%以上,高中低3种浓度的日内和日间RSD〈15%。结论该方法操作简便、灵敏、准确,适用于临床非布司他的血药浓度监测及其药动学研究。

Objective To establish a LC-MS-MS method for determining febuxostate in human plasma. Methods Febuxostate added into blank plasma was sedimented by acetonitrile,and the supernatant was determined by LC-MS-MS. Analytical column was Thermo Biobasic-8,5 μm,50 mm × 2. 1 mm（ ID）. The mobile phase consisted of acetonitrile-10 mmol /L ammonium acetate（ 0. 05%acid = 70∶30 at a flow rate of 0. 2 ml /min. Mass spectrum conditions： ESI-was performed in the SRM mode using target ions m /z 315→271（ 10 eV）（ febuxostate）,m/z 360→274（ 18 eV）（ bezafibrate）,SP 3 500 kV,SGP 10 Arb,AGP45 Arb,TEM 270℃. Results The calibration curve was linear over the range of 10-8 000 μg /L. The LLOQ of Febuxostate in plasma was 10 μg /L. The extracted recovery was〉 85%. The intra-and inter-day RSD were 〈15%. Conclusion The method was sensitive,simple and accurate to determinate febuxostate plasma concentration and to study pharmacokinetics of febuxostate.