基于UPLC-QTOF-MSE代谢组学方法对银杏叶提取物中黄酮醇苷、萜类内酯、酚类及羧酸类成分含量差异研究

摘要

本文建立了超高效液相色谱与三重四级杆飞行时间质谱联用技术UPLC-QTOF-MSE方法,实现对银杏叶提取物中17个黄酮醇苷、5个萜类内酯、4个酚类及7个羧酸类共33个化学成分同时定量分析.运用所建立的方法分析收集的国内外不同来源的银杏叶提取物样品,并借助多元统计的主成分分析方法(PCA)及偏最小二乘法(OPLS-DA)比较分析组分差异,筛选特征成分.结果显示白果内酯、原儿茶酸、莽草酸、奎尼酸、银杏内酯B、银杏内酯J、山柰素-3-O-芸香糖苷、异鼠李素-3-O-芸香糖苷、槲皮素-3-O-α-L-鼠李糖基-2”-(6'''-对香豆酰基)-β-D-葡萄糖苷及芦丁为造成质量差异的特征成分,也被认为是控制银杏叶提取物质量的指标成分.本文建立的方法揭示了银杏叶提取物中不同种类成分原型化合物的准确含量,对其进行更为全面的质量控制;同时通过对特征成分的有效控制,可以保证银杏叶提取物质量均一、稳定,并为其标准化研究提供有力的依据.%In the present study,we established an ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-QTOF-MSE) method to simultaneously quantify 33 components in Ginkgo biloba leaf extracts (GBEs),including 17 flavonol glycosides,five terpene trilactones (TTLs),four polyphenols and seven carboxylic acids.This optimized method was successfully applied to analyze the explicit compositions of GBE samples collected from different places.Furthermore,the data were processed through unsupervised principal component analysis (PCA) and supervised orthogonal partial least squared discrimination analysis (OPLS-DA) to evaluate the quality and compare the differences between the samples according to the contents of the 33 chemical constituents.Bilobalide,protocatechuic acid,shikimic acid,quinic acid,ginkgolide B,ginkgolide J,kaempferol-3-O-rutinoside,isorhamnetin-3-O-rutinoside,quercetin-3-O-α-L-rhamnopyranocyl-2"-(6"-p-coumaroyl)-β-D-glucoside and rutin were recognized as characteristic chemical markers that contributed most to control the quality of GBEs.Based on the fact that GBEs should be standardized with the characteristic components as quality control chemical markers,it is most important to maintain the quality of GBEs stable and reliable,and this method also provided a good strategy to further rectify and standardize the GBEs market.