UPLC-MS/MS法测定复方黄芩片中黄芩苷在大鼠血浆中的浓度

Determination of baicalin by UPLC-MS/MS in rats plasma administered with Fufang Huangqin tablets

目的：建立UPLC-MS/MS方法测定复方黄芩片中黄芩苷在大鼠血浆中的浓度。方法：建立黄芩苷含量测定的UPLC-MS/MS外标分析方法。色谱柱采用C18色谱柱,流动相采用含0.1%醋酸的水-含0.1%醋酸的乙腈,梯度洗脱,在电喷雾电离（ESI）正离子模式下,采用多重反应监测模式（MRM）进行检测。采用清洁级雄性SD大鼠8只,7周龄,体重200±20g,给予复方黄芩片灌胃后（相当于黄芩苷含量200mg/kg）于0.17、0.33、0.5、1、2、4、6、8、10、12、16、24h时间点用断尾法取血0.4 mL,置于肝素化的离心管中,5000r/min离心10min,分离提取血浆,采用UPLC-MS/MS法测定血浆中黄芩苷的含量。结果：血药浓度测定结果表明,黄芩苷的线性范围为5~2000μg/L,定量下限为5μg/L。低、中、高3种浓度的日内和日间精密度相对标准偏差（RSD）不大于3%,平均回收率均在80%以上。黄芩苷血药浓度较低,血药浓度-时间曲线表现为多峰现象。结论：实验建立的UPLC-MS/MS方法符合生物样品分析的要求,可用于研究大鼠灌胃复方黄芩片后血浆中黄芩苷的测定。复方黄芩片口服后其主要药理成分黄芩苷的血药浓度较低,提示药物的吸收利用较差。

Objective：Develop a UPLC-MS/MS method to determinating the plasma concentration of baicalin in Fufang Huangqin tablets.Methods：Develop a UPLC-MS/MS analysis method to determining the baicalin.Using C18 column,mobile phase uses water containing 0.1% acetic acid-acetonitrile containing 0.1% acetic acid,and use ESI positive ion mode and MRM mode for detection.Using 8 male SD rats,7weeks old,weighing 200±20g.Collect 0.4 mL blood from tails at 5,10,20,30,60,120,240,360,480,600,720,960,1440 min after giving Fufang Huangqin tablets orally（Equivalent Baicalin 200mg/kg）,and place it in heparinized centrifuge tube,5000r/min centrifuge 10 min,separate and extract the plasma for UPLC-MS/MS method to determining plasma concentration of baicalin.Results：The results show that the plasma concentration of baicalin has linear range of 5~2000μg/L,the detection limit was 5μg/L.Inter-day and intra-day precision RSD of Low,medium,and high concentrations were not more than 3%,the average recovery was above 80%.The blood concentrations of baicalin was low,plasma concentration-time curve shows a multimodal phenomenon.Conclusion：The UPLC-MS/MS method can be successfully used for measuring the concentration of baicalin in plasma of rats.The blood concentrations of baicalin is low,indicating poor absorption and utilization of drugs.