HPLC-MS/MS测定大鼠血浆中刺五加、贯叶连翘提取物主要活性成分浓度及其药动学研究

HPLC-MS/MS Method for Determination of The Concentration of Main Active Ingredients in The Extracts of Acanthopanax senticosus and Hypericum perforatum L.and Their Pharmacokinetics in Rats

目的建立HPLC-MS/MS测定刺五加提取物、贯叶连翘提取物主要活性成分在大鼠血浆中药物浓度的方法,应用于刺五加提取物、贯叶连翘提取物及其联用时主要活性成分在大鼠体内的药动学研究,并使用DAS 3.2.8软件对主要活性成分的药动学参数进行非房室模型拟合。方法以Welch Ultimate XB-C18(2.1 mm×100 mm,4.6μm)为色谱柱,乙腈(含0.1%甲酸)-0.1%甲酸水溶液(含5 mmol·L^(-1)乙酸铵)为流动相,以300μL·min^(-1)的流速进行梯度洗脱,采用电喷雾离子源进行正负离子同时扫描,多反应监测模式进行监测。结果所建立的方法精密度、准确度良好,提取回收率基本符合生物样品分析方法要求。结论药动学结果表明,相比于单独给药,联合用药后刺五加苷E tmax缩短,金丝桃素ρmax升高、t1/2延长。该药动学结果表明,刺五加提取物、贯叶连翘提取物联用时,很可能通过增加金丝桃素血药浓度、缩短刺五加苷E的达峰时间,从而在体内产生协同抗抑郁作用。

OBJECTIVE To establish a HPLC-MS/MS method for quantitative determination of the main active ingredients of Acanthopanax senticosus extracts and Hypericum perforatum L.extracts in rat plasma,and study combination pharmacokinetics of main active ingredients in rats.METHODS The chromatographic separation was performed on Welch Ultimate XB-C18(2.1 mm×100 mm,4.6μm)column as the mobile phase was acetonitrile(containing 0.1%formic acid)-0.1%formic acid in water(containing 5 mmol·L^(-1)ammonium acetate).The flow rate of 300μL·min^(-1)was used for gradient elution,and the electrospray ion source was used for simultaneous scanning of positive and negative ions,and the multi-reaction monitoring mode was used for detection.RESULTS The results show that the established method had good precision and accuracy,and the extraction recovery rate was basically in line with the requirements of biological sample analysis methods.CONCLUSION Pharmacokinetic results show that compared with single administration,eleutheroside E tmax is shortened,hypericinρmax is increased,and t1/2 is prolonged.The pharmacokinetic results indicat that when the extract of Acanthopanax senticosus is combined with the extract of Hypericum perforatum L.,it might produce synergistic anti-depression effect in vivo by increasing the blood concentration of hypericin and shortening the peak time of eleutheroside E.