UHPLC-MS/MS同时测定清感童饮中6种有效成分在大鼠体内血药浓度及药代动力学研究

Simultaneous determination of six active ingredient in Qinggan Tongyin by UHPLC-MS/MS and pharmacokinetics in rats

关键词:  清感童饮  药代动力学  UHPLC-MS/MS  血浆

Key words:  Qinggan Tongyin  pharmacokinetics  UHPLC-MS/MS  plasma

摘要:

[目的] 本实验旨在研究清感童饮活性成分在大鼠血浆中的药代动力学特征。[方法] 实验建立大鼠血浆中甘草次酸、木犀草素、芹菜素、芍药苷、连翘酯苷A、连翘苷等6种活性成分的超高效液相色谱-串联质谱（UHPLC-MS/MS）检测方法，考察大鼠灌胃给药清感童饮后，6种成分在大鼠体内的药代动力学特征。采用WELCN Ultimate XB-C₁₈色谱柱（2.1 mm×100 mm，3 μm）分离；流动相为0.1%甲酸水（A）-乙腈（B）；梯度洗脱；进样量3 μL；流速为0.2 mL/min；柱温40 ℃；在多反应监测模式（MRM）下正、负离子同时测定。[结果] 结果显示，大鼠血浆样品中各成分的线性关系均良好（r>0.99），其日内、日间精密度、准确度、稳定性、基质效应以及提取回收率均符合生物样品的分析要求。药动学参数结果显示，各成分的t_1/2z为1.18~34.12 h，T_max为0.44~19.33 h，C_max为9.00~318.09 μg/L，AUC_0~t为12.40~15 969.32 h·μg/L，MRT_0~t为2.23~23.19 h。[结论] 本实验所建立的检测方法快速、准确、重复性好，可适用于清感童饮中甘草次酸、木犀草素、芹菜素、芍药苷、连翘酯苷A、连翘苷在大鼠血浆中的药动学研究。

Abstract:

[Objective] The aim of this experiment is to study the pharmacokinetic characteristics of the active ingredients of Qinggan Tongyin in rat plasma. [Methods] An ultra performance liquid chromatography tandem mass spectrometry（UHPLC-MS/MS） method was established to detect six active ingredients in rat plasma，including glycyrrhetinci acid，luteolin，apigenin，paeoniflorin，forsythiaside A，and phillyrin and investigate the pharmacokinetic characteristics of six components in rats after oral administration of Qinggan Tongyin. Using WELCN Ultimate XB-C₁₈ column（2.1 mm×100 mm，3 μm） Separation；The mobile phase is 0.1% formic acid water（A）-acetonitrile（B）；Gradient elution；Injection volume 3 μL；The flow rate is 0.2 mL/min；column temperature is 40 ℃；Simultaneous determination of positive and negative ions in multi reaction monitoring mode（MRM）. [Results] The research results showed that the linear relationships of various components in rat plasma samples were good（r>0.99），and its intra-day and inter-day precision，accuracy，stability，matrix effect and extraction recovery rate all met the analysis requirements of biological samples. The pharmacokinetic parameter results show that the t_1/2z of each component is 1.18 to 34.12 h，T_max is 0.44 to 19.33 h，C_max is 9.00 to 318.09 μg/L，AUC_0~t is 12.40 to 15 969.32（h·μg）/L，and MRT_0~t is 2.23 to 23.19 h. [Conclusion] The detection method established in this experiment is fast，accurate，and has good repeatability，and can be applied to the pharmacokinetic study of glycyrrhetinci acid，luteolin，apigenin，paeoniflorin，forsythiaside A，and phillyrin in rat plasma.