雪莲果叶UPLC-ECD指纹图谱及其抗氧化活性成分研究

Study on UPLC-ECD Fingerprints and Antioxidant Components of Smallanthus Sonchifolius Leaves

目的建立雪莲果叶超高效液相色谱-电化学检测(UPLC-ECD)指纹图谱,并研究其抗氧化成分。方法采用UPLC-ECD法建立10批雪莲果叶样品的指纹图谱,通过《中药色谱指纹图谱相似度评价系统》(2012版)确定共有峰并评价相似度,结合超高效液相色谱-质谱联用(UPLC-MS)技术指认共有峰成分。以DPPH自由基、ABTS^(+)˙自由基清除实验及Fe^(3+)还原能力实验评价10批样品的抗氧化能力,采用灰色关联度法建立谱效关系。结果10批样品共有12个共有峰,指认出6个成分,分别为芦丁、异槲皮苷、异绿原酸A、咖啡酰葡萄糖二酸、双咖啡酰太洛黏酸、2,3,5-或2,4,5-三咖啡酰太洛黏酸。样品相似度在0.983~0.754之间,聚类分析显示10批样品可聚为两类,S5、S7为一类,其余为第二类。灰色关联结果显示,12个共有峰均与抗氧化活性有关联性。结论ECD能有效筛选雪莲果叶中的抗氧化成分,所建雪莲果叶指纹图谱的共有峰均具有抗氧化能力。

Objective To establish the fingerprints of ultra-high performance liquid chromatography coupled to electrochemical detection(UPLC-ECD)of Smallanthus Sonchifolius leaves and analyze the antioxidant components of Smallanthus Sonchifolius leaves.Methods UPLC-ECD was conducted to establish the fingerprints of 10 batches of Smallanthus Sonchifolius leaves and the TCM Chromatographic Fingerprint Similarity Evaluation System(2012 Edition)were used to identify the common peaks and evaluate their similarity of the samples.UPLC-ECD fingerprints combined with UPLC-MS were adopted to identify the common peak components of Smallanthus Sonchifolius leaves.DPPH free radical and ABTS^(+)˙free radical scavenging method and FRAP method were used to evaluate the antioxidant activity of 10 samples and then grey correlation method was used to study the spectrum-effect relationship.Results There were 12 common peaks in the UPLC-ECD fingerprints of 10 samples and 6 components were identified as rutin,isoquercitrin,isochlorogenic acid A,caffeoylglucaric acid and dicaffeoyltaric acid,2,3,5-or 2,4,5-tricaffeoyltaric acid.The similarities of the samples were ranged from 0.983 to 0.754.Cluster analysis showed that 10 batches of samples could be classified into two categories that S5 and S7 as one category and the others as the second category.The results of grey correlation showed that all the 12 common peaks were related to antioxidant activity.Conclusion ECD can effectively screen the antioxidant components in Smallanthus Sonchifolius leaves,and all the common peaks of the fingerprints have antioxidant activity.