木芙蓉叶抗氧化活性成分谱效关系研究

Relationship Between Antioxidant Activity and Spectrum-effect of Hibisci Mutabilis Folium

关键词:  木芙蓉叶  乙酸乙酯部位  抗氧化  指纹图谱  谱效关系

Key words:  Hibisci Mutabilis Folium  ethyl acetate extract  antioxidant  fingerprint  spectrum-activity relationship

摘要:

目的 比较木芙蓉叶不同极性部位的抗氧化活性，测定乙酸乙酯部位抗氧化活性并建立HPLC指纹图谱，研究谱效关系。方法 以DPPH法和ABTS法评价抗氧化活性，筛选木芙蓉叶抗氧化活性最佳部位，测定13批不同产地木芙蓉叶最佳部位的抗氧化活性，并建立HPLC指纹图谱，进行相似度分析、聚类热图分析以及主成分分析(principal component analysis，PCA)，并通过偏最小二乘回归法(partial least squares regression，PLSR)分析其谱效关系。结果 木芙蓉叶不同极性部位抗氧化活性强弱依次为乙酸乙酯部位>正丁醇部位>水部位>石油醚部位。13批木芙蓉叶乙酸乙酯部位有17个共有峰，共指认出5个成分，峰4为秦皮素，峰6为芦丁，峰8为异槲皮苷，峰10为山奈酚-3-O-芸香糖苷和峰15为银椴苷，13批样品的相似度在0.912~0.995。聚类热图分析和PCA将13批木芙蓉叶样品聚为两类；PLSR分析表明，峰3、峰4(秦皮素)、峰6(芦丁)、峰10(山奈酚-3-O-芸香糖苷)和峰12回归系数与抗氧化活性呈正相关，且贡献度较大(VIP>1)，为抗氧化活性的主要有效成分。结论 木芙蓉叶乙酸乙酯部位具有较好的抗氧化活性，木芙蓉叶抗氧化活性为多个成分协同作用的结果，揭示了木芙蓉叶抗氧化活性的药效物质基础。

Abstract:

OBJECTIVE To compare the antioxidant activities of different polar extracts of Hibisci Mutabilis Folium,to determine the antioxidant activity of ethyl acetate extract and to establish HPLC fingerprint in order to study the spectrum effect relationship.METHODS The antioxidant activity was evaluated by DPPH method and ABTS method to screen the best extracts of Hibisci Mutabilis Folium,determine the antioxidant activity of the best extract of 13 batches of Hibisci Mutabilis Folium from different habitats,and establish HPLC fingerprint,for similarity analysis,cluster heat map analysis and principal component analysis (PCA),and the spectral effect relationship was analyzed by partial least squares regression (PLSR).RESULTS The antioxidant activity of different polar extracts of Hibisci Mutabilis Folium was ethyl acetate extract>n-butanol extract>water extract>petroleum ether extract.There were 17 common peaks in the ethyl acetate extract of 13 batches of Hibisci Mutabilis Folium,including 5 components.Peak 4 was fraxetin,peak 6 was rutin,peak 8 was isoquercetin,peak 10 was kaempferol-3-O-rutoside and peak 15 was tilioside,13 samples with a similarity of 0.912-0.995.Cluster heat map analysis and PCA grouped 13 batches of Hibisci Mutabilis Folium samples into two categories;PLSR analysis showed that the regression coefficients of peak 3,peak 4(aesculin),peak 6(rutin),peak 10(kaempferol-3-O-rutoside) and peak 12 were positively correlated with antioxidant activity,and the contribution was large (VIP>1),which was the main effective component of antioxidant activity.CONCLUSION The ethyl acetate extract of Hibisci Mutabilis Folium has good antioxidant activity.The antioxidant activity of Hibisci Mutabilis Folium is the result of the synergistic action of multiple components,which reveals the pharmacodynamic material basis of the antioxidant activity of Hibisci Mutabilis Folium.