UPLC-MS/MS同时测定舒肝宁注射液在大鼠血浆中4种活性成分的含量及其药代动力学研究

Simultaneous determination of four components of Shuganning Injection in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study

该研究旨在揭示舒肝宁注射液在正常大鼠体内的药代动力学特征。实验采用超高液相色谱-电喷雾-串联质谱(UPLC-ESI-MS/MS)技术建立大鼠血浆样品中同时测定绿原酸、栀子苷、千层纸素A及黄芩苷4个活性成分的分析方法,并使用WinN onL in 6.4软件中的非房室模型(NCA)进行药代动力学参数的拟合。经方法学验证,大鼠血浆样品中各成分的线性关系良好(r>0.995),低、中、高剂量的血浆样品的提取回收率和基质效应在79.14%~101.4%。日内、日间精密度、准确度和稳定性均符合生物样品分析要求。药代动力学参数结果表明,绿原酸、栀子苷、千层纸素A在实验剂量下的半衰期(t1/2)没有发生显著性变化,药时曲线下面积(AUC0-t)与剂量成正比,提示3种成分在体内表现出线性动力学的特征,而黄芩苷表现出非线性动力学特征,且各成分在大鼠体内的滞留时间短。建立的UPLC-MS/MS定量分析方法快速、灵敏、准确,可用于大鼠血浆中绿原酸、栀子苷、千层纸素A以及黄芩苷的测定及舒肝宁注射液的药代动力学研究。

This study aims to reveal the pharmacokinetics of Shuganning Injection in normal rats.In this experiment,ultra-high performance liquid chromatography-electrospray-tandem mass spectrometry(UPLC-ESI-MS/MS)was used to establish an analytical method for simultaneous determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma.Then,the non-compartmental model(NCA)in Phoenix WinN onL in 6.4 software was used to fit pharmacokinetic parameters.The methodological validation showed that the linear relationship of the components in rat plasma samples were good(r>0.995).The recovery rate and matrix effect of plasma samples with low,middle and high concentration were 79.14%-101.4%.The intra-day and inter-day precision,accuracy and stability meet the requirements of biological sample analysis.The half-life(t1/2)of chlorogenic acid,gardenioside,oroxylin A did not change significantly and the area under blood concentration-time curve(AUC0-t)is proportional to the dose,which suggested that three components showed a linear kinetic characteristics,but baicalin showed nonlinear kinetic characteristics.Moreover,the retention time of each component in rats was short.The established UPLC-MS/MS quantitative analysis method is rapid,sensitive and accurate,which can be used for the determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma and pharmacokinetic study of Shuganning Injection.