基于高效液相色谱-串联质谱和分子网络联合应用技术快速鉴定川黄柏和关黄柏中的化学成分

Rapid identification of components in Phellodendri Chinensis Rupr and Phellodendri Amurensis Schneid with HPLC-MS and molecular network

目的通过高效液相色谱-串联质谱和分子网络整合技术联合应用的快速分析方法分别鉴定川黄柏与关黄柏中的化学成分。方法采用Agilent C18色谱柱(100 mm×4.5 mm,1.8μm),流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱。通过三重四极杆-飞行时间串联质谱检测数据,根据MS/MS碎片模式的相似性创建分子网络,在正离子模式下,将拥有相似结构的分子簇运用MSDIAL软件筛选出来。最后结合保留时间信息、高分辨质谱精确相对分子质量和MS/MS碎片信息,并根据相关文献和分子网络进行比对,分别对川黄柏、关黄柏中化学成分进行快速准确鉴定。结果在川黄柏、关黄柏中共鉴定出22种化学成分,并通过质谱的积分面积确定了两者的相对峰面积比例为2.08∶1。通过分子网络分析发现川黄柏中存在很多响应较低的未被鉴定出的生物碱类成分,而木兰花碱同系物多出现于关黄柏中,并且关黄柏中的巴马汀的离子响应较川黄柏更高。结论该方法可为关黄柏、川黄柏的质量控制、基准物质以及药效物质研究提供参考,同时也可用于两者化学成分的快速定性分析。

Objective To rapidly identify the chemical compositions in Phellodendri Chinensis Rupr(PC)and Phellodendri Amurensis Schneid(PA)with high performance liquid chromatographytandem mass spectrometry(HPLC-MS)and molecular network.Methods Agilent C18 column(100 mm×4.5 mm,1.8μm)was used with 0.1%formic acid water(A)-acetonitrile(B)as the mobile phase for gradient elution.Detection data by quadrupole-time of flight-mass spectrometry were collected in the positive ion mode,and molecular network was created according to the similarity of MS/MS fragmentation modes.MSDIAL software was used to screen molecular clusters with similar structures.Finally,the chemical compositions of PC and PA were confirmed according to the retention time,accurate molecular weight of high resolution mass spectrometry and MS/MS multi-stage fragments.Results Totally 22 chemical constituents were identified in PC and PA.The relative proportion of alkaloids in PC and PA was 2.08∶1 by integral area of mass spectrum.Molecular network found many unknown alkaloids with low response in PC.There were more magnoflorine homologues in PA,and the ion response of palmatine in PA was higher than that in PC.Conclusion This method can provide reference for the quality control and pharmacodynamic substances of PC and PA,and rapid qualitative analysis of chemical components of traditional Chinese medicine.