HPLC-MS/MS法测定人血浆中利伐沙班的浓度  

Determination of rivaroxaban concentration in human plasma by HPLC-MS/MS

作　　者：陈敏[1] 高红瑾[1] 阳丽梅[1] CHEN Min;GAO Hongjin;YANG Limei(Department of Pharmacy,Fujian Provincial Hospital,Provincial Clinical Medical College of Fujian Medical University,Fuzhou,Fujian 350001,China)

机构地区：[1]福建医科大学省立临床医学院福建省立医院药学部

出　　处：《福建医药杂志》2019年第6期145-148,共4页Fujian Medical Journal

摘　　要：目的建立人血浆中利伐沙班(rivaroxaban)的高效液相色谱-串联质谱法(HPLC-MS/MS)法测定的方法,并对其进行方法学验证。方法血浆样品采用乙腈直接沉淀,选择盐酸氨溴索作为内标物。采用菲罗门Kinetex 2.6u C18100A(50×2.10 mm,1.8μm)色谱柱,流动相为乙腈∶水(30∶70)用甲酸调pH至3.0,流速0.3 mL/min,柱温35℃,电喷雾离子源(ESI)正离子检测方式,选择多重反应监测模式进行测定,用于定量的离子对分别为利伐沙班质荷比(m/z)436.0/144.9、盐酸氨溴索m/z 379.0/263.9。结果利伐沙班在2.5~500 ng/mL范围内呈良好的线性关系(r=0.9999),定量下限是2.5 ng/mL;日内、日间相对标准偏差(RSD)0.29%~2.38%,相对误差(RE)―0.59%~0.90%,基质效应97.76%~100.41%,提取回收率为98.64%~102.84%。血浆样品在室温放置24 h、―20℃反复冻融3次以及―80℃冷冻30 d,不影响利伐沙班含量测定。结论本研究建立的HPLC-MS/MS法测定利伐沙班血药浓度,具有简便、快速、准确,适用于临床常规监测及药动学研究等特点。Objective To establish a HPLC-MS/MS method to determinate rivaroxaban concentration in human plasma,and the method was verified by methodology.Methods Plasma samples by acetonitrile precipitation were to be determined by using ambroxol hydrochloride as Internal Standard(IS).The chromatographic column was Kinetex 2.6u C18100A(50×2.10 mm,1.8μm),and the mobile phase using water(adjust pH to 3.0 with formic acid)-acetonitrile(70∶30)with flow rate of 0.3 mL/min.The mass spectrometer was operated in positive electron spray lonization(ESI)and multiple response monitoring patterns,m/z 436.0/144.9 and m/z 379.0/263.9 were used for the measurement of rivaroxaban and ambroxol hydrochloride,respectively.Results Rivaroxaban was linear in the range of 2.5 to 500 ng/mL,and the quantification lower limit of rivaroxaban was 2.5 ng/mL.The intra-day and inter-day precisions(relative standard deviation,RSD)were 0.29%to 0.38%,the accuracy was between―0.59% and 0.90%.The matrix effect was in the range of 97.76%to 100.41%,and the absolute recovery was in the range of 98.64% to 102.84%.The plasma samples were stable under conditions including placing at room temperature for 24 h,repeated freezing and thawing at―20℃ for 3 times,and freezing at―80℃ for 30 days.Conclusion The HPLC-MS/MS method established in our study is convenience,accurate and rapid for the determination of rivaroxaban in blood,which is applicable for the routine monitoring and PK study of rivaroxaban.

关 键 词：利伐沙班 HPLC-MS/MS法 血药浓度 

分 类 号：R917[医药卫生—药物分析学]