UPLC-MS/MS方法测定健康人体氟氯西林血浆药物浓度及其药动学研究

Determination of Flucloxacillin in Human Plasma by Ultra Performance Liquid Chromatography-tandem Mass Spectrometry and Subsequent Application to a Clinical Study in Healthy Chinese Volunteers

目的：应用超高效液相串联质谱联用方法（UPLC-MS/MS）测定健康人体氟氯西林血浆药物浓度,并对其药动学特征进行分析。方法：采用开放、随机设计试验方案,选择24位健康志愿者,男女各半,随机分成3组,分别静脉给予1.0 g、2.0 g、4.0 g三个剂量试验药物,采用UPLC-MS/MS方法测定血浆中氟氯西林药物浓度。流动相由乙腈：10 mmol·L^-1甲酸铵水溶液=35∶65（v/v）组成。色谱柱为BEHTM C18（Waters）,1.7μm,柱长100 mm×2.1 mm I.D,流速为0.2 mL·min-1,柱温40℃,自动进样器温度4℃,进样量2.5μL。质谱条件：离子源温度120℃;去溶剂气温度：350℃;氟氯西林（m/z 454.1〉160.3）,内标氯唑西林钠（m/z 436.1〉277.3）。结果：氟氯西林标准曲线线性范围为0.2μg·mL^-1~1000μg·mL^-1,线性关系良好（r〉0.99）,最低定量限：0.2μg·mL^-1,高、中、低3种浓度的批内和批间变异（RSD%）均小于15.0%,血浆提取回收率为89.7%~114.7%。结论：本方法处理过程简单可靠,可快捷、准确地应用于氟氯西林的血浆药物浓度测定。

Objective： To establish an UPLC-MS/MS method for determination of flucloxacillin in human plasma and study its pharmacokinetics in healthy subjects. Methods： Twenty-four healthy volunteers were randomized into three dose groups and received 1.0 g,2.0 g or 4.0 g flucloxacillin by intravenous injection. The UPLC method was performed on a BEH C18 OSD column（100 mm×2.1 mm,1.7 μm） with a mixture of 65% 10 mmol·L^-1ammonium formate（NH4 COOH） and 35% acetonitrile as mobile phase. The flow rate was 0.2 mL·min-1and the column temperature was 40℃. The analyses were carried out by multiple reaction monitoring（MRM） using the precursor-to-product combinations of m/z 454.1 →160.3（flucloxacillin） and m/z 436.1→277.3（IS）. Results： The linear range of flucloxacillin was 0.2 to 1000 μg·mL^-1（r0.99）. The LLOQ was 0.2 μg·mL^-1. The precision of intro-batch（RSD%） and inter-batch（RSD%） were less than 15.0%（n=5）,the mean recovery was between 89.7%~114.7%. Conclusion： The method is simple,convenient and accurate,and can be used for detecting flucloxacillin concentration in human plasma.