高效液相色谱串联质谱同时测定柴胡配方颗粒中柴胡皂苷a、b_1、b_2、c的含量

Simultaneous Determination of Saikosaponin a,b_1,b_2 and c in Radix Bupleuri Formula Granules by HPLC-MS/MS

目的建立高效液相色谱串联质谱同时测定柴胡配方颗粒中柴胡皂苷a、b_1、b_2、c的含量,为其质量标准的研究提供依据。方法采用Kinetex C_(18)色谱柱(2.1 mm×100 mm,2.6μm);流动相为乙腈-0.1%甲酸水溶液,梯度洗脱;流速为300μL·min^(-1);柱温40℃;质谱条件:采用电喷雾离子源负离子模式进行检测,多反应监测模式(MRM)用于定量测定,源喷射电压为-4 500 V,离子源温度为550℃。结果柴胡皂苷a、b_1、b_2、c的峰面积与质量浓度在测定范围内均呈良好的线性关系,r≥0.999 1;平均加样回收率分别为100.56%、98.50%、97.44%、99.12%。结论该方法同时测定了柴胡配方颗粒中柴胡皂苷a、b_1、b_2、c的含量,方法可行,重现性好,可为柴胡配方颗粒的质量控制提供方法参考。

OBJECTIVE To establish a method for determining the contents of saikosaponin a, b1, b2 and c in Radix Bupleuri formula granules by HPLC-MS/MS, and provide scientific basis for the quality standards. METHODS The samples were separated on a Kinetex Ct8 column （2. 1 mm× 100 mm, 2. 6μm） by gradient elution at the flow rate of 300μL·min-1 using acetonitrile and 0. 1% aqueous formic acid as the mobile phase. The column temperature was maitained at 40℃. Muhiple reaction monitoring scan- ning （MRM） was employed for the quantification with switching electrospray ion source polarity in negative mode. The ion spray volt- age was set at -4 500 V and the turbo spray temperature was maintained at 550℃. RESULTS There was significant correlation be- tween the peak area and concentration of each compound within the test range （ r ≥ 0. 999 1 ）. The average recoveries were from 97.44% to 100. 56%. CONCLUSION Saikosaponin a, b1 , b2 and c are determined by this method simultaneously. The method is available with good reproducibility and can control the quality of Radix Bupleuri formula granules effectively.