拳参高效液相色谱指纹图谱与抗氧化活性的谱效关系研究

Study on the spectrum effect relationship between high performance liquid chromatography fingerprint and antioxidant activity of Polygonum bistorta L.

目的采用高效液相色谱(HPLC)指纹图谱与抗氧化活性建立谱-效关系分析筛选拳参抗氧化活性的有效成分。方法采用HPLC法建立拳参指纹图谱,评价指纹图谱相似度,结合HPLC-Q-Orbitrap HRMS/MS定性分析,对其中29个主要特征峰进行指认。使用2,2′-联氮-双(3-乙基苯并噻唑啉-6-磺酸)二胺盐(ABTS)和2,2-二苯基-1-苦基肼(DPPH)自由基清除法,以及铁离子还原能力(FRAP)测定法,分别评价拳参样品的抗氧化能力,并利用抗氧化活性综合指数(APC)进行综合评价,结合灰色关联度分析和偏最小二乘回归分析研究谱效关系。结果建立了11批拳参样品的HPLC指纹图谱,相似度评价结果均≥0.970。标定29个主要特征峰,并鉴定其中26种成分。灰色关联度分析和偏最小二乘回归分析结果显示,峰2+3(6-O-没食子酰葡萄糖)、峰8(二没食子酰葡萄糖)、峰11(绿原酸)、峰14(儿茶素)、峰15(二没食子酰葡萄糖)、峰18(二没食子酰葡萄糖)和峰23(表儿茶素)是反映拳参抗氧化药效的重要特征峰。结论拳参发挥的抗氧化作用是酚酸类和黄酮类等多种成分联合效应的结果,本文研究结果可为拳参药材的质量评价提供更全面的参考。

Objective The high performance liquid chromatography(HPLC)fingerprints were correlated with the antioxidant activity to establish a spectrum-effect relationship to screen the active ingredients with the antioxidant activities of Polygonum bistorta L.Methods The fingerprint of P.bistorta L.was established by HPLC and the similarity of the fingerprint was evaluated.Combined with HPLC-Q-Orbitrap HRMS/MS qualitative analysis,29 of the major characteristic peaks were designated.The antioxidant activity of different batches was evaluated by the free radical scavenging rate of 2,2′-diazo-bis(3-ethylbenzothiazolin-6-sulfonic acid)diamine salt(ABTS),2,2-diphenyl-1-trinitrophenylhydrazine(DPPH),and total antioxidant capacity(FRAP).The antioxidant activity comprehensive index(APC)was used for a comprehensive evaluation.Grey relational analysis and partial least square regression analysis were used for a comprehensive evaluation.Results The HPLC fingerprints of 11 samples were established,with the similarity greater than or equal to 0.970,demarcating 29 main characteristic peaks,and identifying 26 components.The results of grey relational analysis and partial least square regression analysis showed that peaks 2+3(6-O-Galloylglucose),8(Digallic glucose),11(Chlorogenic acid),14(Catechin),15(Digallic glucose),18(Digallic glucose),and 23(Epicatechin)were the important characteristic peaks reflecting the antioxidant efficacy of P.bistorta L.Conclusion The antioxidant activity of P.bistorta L.is the result of the combined effect of multi-components such as phenolic acids and flavonoids,which provides a more comprehensive reference for the quality control of medicinal materials.