苗药地瓜藤UHPLC指纹图谱及抗氧化活性谱效关系研究

UHPLC Fingerprinting and Spectroscopic Relationship of Antioxidant Activity of the Miao Medicine Ficus Tikoua Bur.

关键词:  地瓜藤  指纹图谱  抗氧化  灰色关联度  偏最小二乘回归分析  谱效关系

Key words:  Ficus tikoua Bur.  fingerprint  anti-oxidation  gray correlation  partial least squares regression  spectral efficiency relationship

摘要:

目的 建立苗药地瓜藤的UHPLC指纹图谱，研究其与抗氧化活性的谱效关系，筛选抗氧化活性成分群。方法 采用UHPLC建立地瓜藤指纹图谱，运用《中药指纹图谱相似度评价系统》(2012版)进行相似度评价并指认共有峰，运用SPSS 16.0、SIMCA 14.1软件进行聚类分析(hierarchical cluster analysis，HCA) 和主成分分析(principal component analysis，PCA)；采用1,1-二苯基-2-三硝基苯肼自由基清除法、2,2’-联氮-二(3-乙基苯并噻唑啉-6-磺酸)二铵盐自由基清除法和总抗氧化能力测定法评价16批地瓜藤抗氧化活性；运用灰色关联度分析(grey relational analysis，GRA)、双变量相关分析、偏最小二乘回归(partial least squares regression，PLSR)3种分析方法研究谱效关系。结果 建立了16批地瓜藤的UHPLC指纹图谱并标定13个共有峰，相似度为0.613~0.996，同时指认出峰9为芦丁、峰10为异槲皮素、峰12为水仙苷，HCA结果显示样品共聚为两类，与PCA结果一致；16批地瓜藤均有不同程度抗氧化活性。GRA结果显示13个共有峰与抗氧化活性之间的关联度>0.8，均具有高关联性，双变量相关性分析及PLSR分析结果表明峰5、峰9(芦丁)、峰10(异槲皮素)、峰11、峰12(水仙苷)的相关系数及回归系数与抗氧化活性呈正相关，且贡献度较大(变量重要性投影>1)，为抗氧化活性的主要有效成分。结论 16批地瓜藤均具有较好的抗氧化活性，其发挥抗氧化作用是内部抗氧化成分群协同作用的结果，而共有峰5，9，10，11，12所对应的成分与其抗氧化活性密切相关，揭示了地瓜藤抗氧化活性的药效物质基础。

Abstract:

OBJECTIVE To establish the UHPLC fingerprint of Miao Medicine Ficus tikoua Bur., study its spectrum-effect relationship with antioxidant activity, and screen the antioxidant active components. METHODS UHPLC was used to establish the fingerprint of Ficus tikoua Bur.. Evaluation System for Similarity of Chromatographic Fingerprint of Chinese Herbal Medicine (Version 2012) was used to evaluate the similarity and identify the common peaks. SPSS 16.0 and SIMCA 14.1 software were used for hierarchical cluster analysis(HCA) and principal component analysis(PCA). The 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical scavenging method, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt(ABTS) free radical scavenging method and total antioxidant capacity method were used to evaluate the antioxidant activity of 16 batches of ethanol extracts from Ficus tikoua Bur. Three methods including grey relational analysis(GRA), bivariate correlation analysis and partial least squares regression(PLSR) were used to study the spectrum-effect relationship. RESULTS The UHPLC fingerprints of 16 batches of Ficus tikoua Bur. were established and 13 common peaks were identified. The similarities were 0.613-0.996. At the same time, it was identified that peak 9 was rutin, peak 10 was isoquercetin, and peak 12 was narcissin. The results of HCA showed that the samples were clustered into two categories, which was consistent with the PCA results. Sixteen batches of Ficus tikoua Bur. had different degrees of antioxidant activity. The results of GRA showed that the correlation between 13 common peaks and antioxidant activity was >0.8, and all of them had high correlation. The results of bivariate correlation analysis and PLSR analysis showed that the correlation coefficient and regression coefficient of peak 5, peak 9(rutin), peak 10(isoquercetin), peak 11 and peak 12(narcissin) were positively correlated with antioxidant activity, and the contribution rate was larger(variable importance in projection>1), which were the main active components of antioxidant activity. CONCLUSION All the 16 batches of Ficus tikoua Bur. have good antioxidant activity, and its antioxidant effect is the result of the synergistic action of the internal antioxidant component group. The components corresponding to the common peaks 5, 9, 10, 11, 12 are closely related to their antioxidant activity, revealing the pharmacodynamic material basis of the antioxidant activity of Ficus tikoua Bur.