用LC-MS/MS法同时测定人血浆中甘草酸及其活性代谢产物甘草次酸的浓度  

Simutaneous determination of glycyrrhizin and its active metabolite glycyrrhetinic acid in human plasma by LC-MS/MS

作　　者：黄辉 邓远雄[1] 刘海艳 吴慧慧 HUANG Hui;DENG Yuan-xiong;LIU Hai-yan;WU Hui-hui(College of Medicine,Hunan Normal University,Changsha 410013,Hunan Province,China;Changsha King--eagle Med Technology Co.,Ltd,Changsha 410205,Hunan Province,China)

机构地区：[1]湖南师范大学医学院,湖南长沙410013 [2]长沙晶易医药科技股份有限公司,湖南长沙410205

出　　处：《中国临床药理学杂志》2024年第9期1336-1340,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的建立LC-MS/MS法同时测定人血浆中甘草酸(GLY)及其主要活性代谢产物甘草次酸(GA)的浓度。方法人血浆用甲醇沉淀蛋白后,用Synergi TM 4μm Fusion-RP 80A色谱柱(2.0 mm×50 mm,4μm)进行分离,流动相为乙腈(含0.1%甲酸)-水(含0.1%甲酸),梯度洗脱,流速为0.6 mL·min^(-1)。用电喷雾离子源,负离子模式,多反应监测(MRM)扫描方式进行监测。考察该方法的专属性、线性范围、精密度与准确度、提取回收率、基质效应、稀释验证和稳定性等。结果甘草酸和甘草次酸分别在0.30~80.00 ng·mL^(-1)和1.80~480.00 ng·mL^(-1)线性关系良好,定量下限分别为0.30和1.80 ng·mL^(-1),提取回收率分别为98.14%~109.10%和97.25%~100.87%,甘草酸和甘草次酸都没有明显的基质效应。质控样品批内和批间精密度均小于10.72%,准确度在95.38%~109.34%。结论该方法快速、准确,灵敏度高、专属性强,可用于甘草酸及其活性代谢产物甘草次酸的人体血药浓度检测及复方甘草酸苷片的药代动力学研究。Objective To develop an LC-MS/MS method for the simutaneous determination of glycyrrhizin(GLY)and its main active metabolite glycyrrhetinic acid(GA)in human plasma.Methods Sample preparation was done by protein precipitation with methanol.Chromatographic separation was achieved by a Synergi TM 4μm Fusion-RP 80A(2.0 mm×50 mm,4μm)column with a gradient mobile phase consisting of acetonitrile(containing 0.1%formic acid)-water(containing 0.1%formic acid),at a flow rate of 0.6 mL·min^(-1).GLY and GA were monitored using negative electrospray triple quadrupole mass spectrometer via multiple reaction monitoring(MRM)mode.The specificity,calibration curve and range,accuracy and precision,recovery,matrix effect,dilution integrity and stability were validated.Results The validated method had an excellent linearity in the range of 0.30-80.00 ng·mL^(-1)and 1.80-480.00 ng·mL^(-1)for GLY and GA,with the limits of quantification were 0.30 and 1.80 ng·mL^(-1).Recovery efficiencies were in the range of 98.14%-109.10%and 97.25%-100.87%for GLY and GA.No significant matrix effects were found for the two analytes.Intra-day and inter-day precisions of four quality control concentrations were less than 10.72%.Intra-day and inter-day accuracies were determined to be 95.38%-109.34%with all accuracy measurements.Conclusion The validated method was rapid,sensitive,selective and accurate for the quantification of GLY and its active metabolite GA,successfully used to the pharmacokinetics study of GLY and its active metabolite in human.

关 键 词：甘草酸 甘草次酸 液相色谱串联质谱 药代动力学 

分 类 号：R28[医药卫生—中药学]