UPLC-QQQ-MS测定中药材中马兜铃酸Ⅰ的含量

Determination of Aristolochic Acid Ⅰ in Traditional Chinese Medicine by UPLC-QQQ-MS

目的:利用UPLC-QQQ-MS建立复杂中药基质中马兜铃酸Ⅰ的定性定量测定方法。方法:以马兜铃科中药材及其易于混淆的中药材为研究对象,色谱分离使用Agilent ZORBAX XDB-C18色谱柱(2.1 mm×50 mm,3.5μm),流动相0.2%甲酸水溶液-甲醇梯度洗脱,流速0.2 m L·min^(-1),柱温35℃,采用电喷雾离子源(ESI源),正离子模式下,多反应监测模式(MRM)检测,以吲哚美辛为内标计算浓度。结果:马兜铃酸Ⅰ质量浓度在21.65~1 732μg·L^(-1)线性关系良好(r=0.997 2),加样回收率为82.87%~89.22%,平均加样回收率86.07%(n=6),RSD 2.5%,精密度及方法重复性均良好。基于这个方法笔者将从全国各地收集来的中药材进行定向马兜铃酸Ⅰ含量的检测,结果精准可信。结论:该方法简便,快速,灵敏度高,专属性强,可以用于含有或者可能含有马兜铃酸Ⅰ成分中药材的检测。结果表明,不同种类药材中马兜铃酸Ⅰ的含量差异明显,尤以朱砂莲中马兜铃酸Ⅰ含量最高。同时,该检测结果也可为马兜铃科中药材中马兜铃酸Ⅰ的限量及其合理使用提供实验基础和参考依据。

Objective ： To establish a qualitative and quantitative determination method by using ultra high performance liquid chromatography-tandem triple quadrupole mass spectrometry, in order to determine aristolochic acid I in complex traditional Chinese medicine matrix. Method： Chinese medicinal materials of aristolochiaceae and other easily confused species were used as the research object. Chromatographic separation was performed with Agilent ZORBAX XDB-CIs column （2. 1 mm ×50 mm, 3.5 μm） by gradient elution with mobile phase of 0.2% formic acid in water-methanol. The flow rate was 0. 2 mL.min -1 , and the column temperature was 35 ℃. The target compounds were analyzed in the multiple reaction monitoring mode （MRM） with positive electrospray ionization （ESI 1）. The concentration was calculated with indomethacin as the internal standard. Result： Aristolochic acid I showed a good linearity within the range of 21.65-1 732 μg.L-1 （r =0. 997 2）. The recovery was 82. 87% -89.22% , the average recovery rate was 86. 07% （n = 6）, and the RSD was 2. 5%. Precision of the instrument and repeatability of the method were good. Based on this method, the content of aristolochic acid I in the Chinese herbal medicines collected nationwide was determined, and the results were accurate and reliable. Conclusion： This method is simple, quick, highly sensitive and specific, and can be used for the detection of Chinese medicinal materials containing or possibly containing aristolochic acid I component. The results of resource data showed that the contents of aristolochic acid I in different kinds of herbs were significantly different, especially in Aristolochia tuberosa, which had the highest content. The results can also provide an experimental basis and reference for content limit of aristolochic acidIin Aristolochiaceae plants and its reasonable use.