利伐沙班在深静脉血栓形成患者中的药动学研究

Pharmacokinetics Study of Rivaroxaban in Patients Treated for Deep-Vein Thrombosis

目的建立测定人血浆中利伐沙班浓度的UPLC-MS/MS方法,测定深静脉血栓患者体内利伐沙班血药浓度并进行药物动力学研究。方法血浆经乙腈沉淀蛋白,采用Waters Acquity BEH C18色谱柱(2. 1 mm×50 mm,1. 7μm),流动相为乙腈和水,梯度洗脱,流速为0. 4 m L·min-1;采用电喷雾离子化源,正离子模式,MRM进行监测,m/z 437. 3→145. 0(利伐沙班)和m/z 440. 1→145. 0(d4-利伐沙班)。6名深静脉血栓患者首次服用20 mg利伐沙班片,采集血浆样本进行检测。结果血浆中利伐沙班在0. 5～400 ng·m L-1内线性关系良好(r=0. 998 3),定量下限0. 5 ng·m L-1;批内、批间RSD均<15%;提取回收率稳定,无明显基质效应。深静脉血栓患者服用利伐沙班后约2 h达峰,平均峰浓度为317. 4 ng·m L-1,半衰期为6. 5 h。结论所建方法快速准确、专属性强、重复性好,适用于DVT患者血浆中利伐沙班血药浓度的检测和药动学研究。

OBJECTIVE To develop an UPLC-MS/MS method for the determination of rivaroxaban in human plasma and apply it to the pharmacokinetics study. METHODS The plasma samples were precipitated by acetonitrile. The Waters Acquity BEH C18 column（ 2. 1 mm ×50 mm,1. 7 μm） was adopted. The mobile phase was acetonitrile and water with the gradient elution at the flow rate of 0. 40 m L·min^-1. Detection of the analyte was achieved by using positive ion electrospray ionization（ ESI） in the multiple reaction monitoring（ MRM） mode. The MS/MS iontransitions monitored were m/z 437. 3→145. 0 and m/z 440. 1→145. 0 for rivaroxaban and internal standard,respectively. Blood plasma samples were collected and tested after patients with deep-vein thrombosis（ n = 6） took single oral dose of rivaroxaban tablets（ 20 mg）. RESULTS The linear range of rivaroxaban was 0. 5-400 ng·m L^-1（ r = 0. 998 3）. The lower limit of quantitation was 0. 5 ng·m L^-1 and the intra-day and inter-day relative standard deviations were 15%. The recovery rate was stable and nosignificant matrix effect was found. The maximum plasma concentration（ ρmax） of rivaroxaban in patients with deep-vein thrombosis was317. 4 ng·m L^-1,and the time to ρmaxwas 2. 0 h,and the elimination half-life（ t1/2） in plasma was 6. 5 h. CONCLUSION This method is rapid,specific,reliable and suitable for the determination of rivaroxaban in human plasma and pharmacokinetic study.