改进的LC-MS/MS法同时测定人血浆中奥司他韦及其活性代谢产物浓度  

Simultaneous determination of oseltamivir and its active metabolite in human plasma by an improved LC-MS/MS method

作　　者：容志欢 张博 李鑫 相佳宏 石晓伟[1] 李德强[2] RONG Zhi-huan;ZHANG Bo;LI Xin;XIANG Jia-hong;SHI Xiao-wei;LI De-qiang(School of Pharmacy,Hebei Medical University,Shijiazhuang 050017,Hebei Province,China;Department of Pharmacy,The Second Hospital of Hebei Medical University,Shijiazhuang 050051,Hebei Province,China)

机构地区：[1]河北医科大学药学院,河北石家庄050017 [2]河北医科大学第二医院药学部,河北石家庄050051

出　　处：《中国临床药理学杂志》2021年第11期1425-1429,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河北省自然科学基金资助项目(H2020206067)。

摘　　要：目的改进一种同时测定人血浆中奥司他韦及其活性代谢产物奥司他韦酸浓度的LC-MS/MS方法。方法人全血预先加入双-(对-硝基苯基)磷酸酯(BNPP;酯酶抑制药)处理,血浆样本加入甲醇沉淀蛋白。色谱柱为Synergi fusion-RP C18(50.0 mm×3.0 mm, 4μm),流动相为0.2%甲酸-甲醇,梯度洗脱,流速为0.6 mL·min^(-1)。用电喷雾正离子多反应监测。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。结果奥司他韦和奥司他韦酸的线性浓度范围分别为0.4~200.0 ng·mL^(-1)和2.0~1 000.0 ng·mL^(-1),线性相关系数均>0.998 0,日内标准偏差均小于6.24%,日间标准偏差均小于5.23%,绝对回收率可达90.82%以上。样本稳定性良好。结论本方法用BNPP作为酯酶抑制药,改进了LC-MS/MS法同时测定血浆中奥司他韦及其活性代谢产物奥司他韦酸药物浓度,适用于临床治疗药物监测。Objective To improve a LC-MS/MS method for the simultaneous determination oseltamivir and its active metabolite oseltamivir carboxylate in human plasma. Methods Human whole blood was pretreated with the esterase inhibitor Bis(p-nitrophenyl) phosphate(BNPP). The plasma samples were precipitated with methanol. The separation was performed on a Synergi fusion-RP C18(50.0 mm×3.0 mm, 4 μm) using a mobile phase of 0.2% formic acid and methanol with a gradient elution at a flow rate of 0.6 mL·min^(-1). The analytes were detected by multiple reaction monitoring mode in positive electrospray ionization. The specificity, standard curve, lower limit of quantitation, precision, recovery, matrix effect and stability of the method were investigated. Results Oseltamivir and oseltamivir carboxylate were linear in ranges of 0.4-200.0 ng·mL^(-1) and 2.0-1 000.0 ng·mL^(-1) with the coefficients of linearity more than 0.998 0. The intra-day RSD were all less than 6.24%. The inter-day RSD were all less than 5.23%. The absolute recovery was above 90.82%. The stability was good. Conclusion The improved LC-MS/MS method using BNPP as an esterase inhibitor was successful in determining oseltamivir and its active metabolite oseltamivir carboxylate in human plasma that is suitable for clinical therapeutic drug monitoring.

关 键 词：奥司他韦 活性代谢物 治疗药物监测 人血浆 液相色谱-串联质谱 

分 类 号：R978.7[医药卫生—药品]