UPLC-MS/MS法测定清肺抑火丸中4种有效成分的含量

目的：建立UPLC-MS/MS法同时测定清肺抑火丸中知母皂苷BⅡ/BⅢ、菝葜皂苷元和桔梗皂苷元的含量。方法：采用Waters ACQUITY UPLC BEH C₁₈（2.1 mm×100 mm，1.7 μm）色谱柱，以水-乙腈为流动相，流速为0.2 mL·min^-1，柱温为40℃，进样量为2 μL；采用电喷雾离子源在正、负模式下以多反应监测方式进行定量分析；通过样品测定值的分析比较，评价不同批次及不同企业样品间的质量差异。结果：4种成分均在检测范围内进样量与峰面积线性关系良好，方法精密度、稳定性、重复性和回收率均符合要求。比较测定结果发现各企业批间样品均一性较好，但不同企业间样品的测定结果差异显著，仅1家企业的样品符合限度要求。相关性分析结果显示样品中知母皂苷BⅡ/BⅢ的总量与菝葜皂苷元含量呈正相关。结论：该法简便快捷、重复性好、准确度高，可提升清肺抑火丸的质量控制水平。

Determination of four active components in Qingfei Yihuo Pills by UPLC-MS/MS

OBJECTIVE To establish a UPLC-MS/MS method for the determination of timosaponin BⅡ, anemarsaponin BⅢ, sarsasapogenin and platycodigenin in Qingfei Yihuo Pills.METHODS A Waters ACQUITY UPLC BEH C₁₈ column(2.1 mm×100 mm, 1.7 μm) was adopted with water-acetonitrile as the mobile phase.The flow rate was 0.2 mL·min^-1, the column temperature was 40℃and the injection volume was 2 μL.And electrospray ionization(ESI) was utilized for quantitative analysis by multiple reaction monitoring(MRM) in positive and negative modes, respectively.The quality differences in samples from the different enterprises were revealed through analyzing the determination results of samples.RESULTS The four components had an excellent linear relationship between the injection amount and peak area within the determination range.And the precision, stability, repeatability and average recoveries complied with the requirements.The comparison results indicated the uniformity of samples from the same enterprise was good, but the different enterprises showed significant differentiation.Only one enterprise's sample met the limit requirements.Meanwhile, it was found that there was a positive correlation between the total amount of timosaponin BⅡ and anemarsaponin BⅢ and the content of sarsasapogenin in the samples.CONCLUSION The established method is simple, reliable and accurate which can improve the current quality control of Qingfei Yihuo Pills.