目的: 建立甘草药材的HPLC指纹图谱,并研究其与抗氧化活性之间的谱效关系。方法: 采用HPLC法建立甘草的指纹图谱;在与对照图谱相似度评价均大于0.915的基础上,结合聚类分析(hierarchical cluster analysis,HCA)和主成分分析(principal component analysis,PCA),对其中的25个共有峰进行评价;1,1-二苯基-2-三硝基苯肼(DPPH)法,铁离子还原能力(FRAP)法及水杨酸法评价甘草抗氧化活性;灰色关联度分析(grey relational analysis,GRA)和偏最小二乘回归分析(partial least squares regression analysis)研究谱效关系。结果: 建立了甘草的HPLC指纹图谱,确定了其中26个色谱峰为共有峰,相似度均在0.915以上,通过对照品比对法指认了其中9个色谱峰。HCA结果表明,全部样品被聚为4类,与PCA结果基本一致,25批甘草均有不同程度抗氧化活性。谱效关系表明2、7、10、13、18、21、23号峰是与抗氧化活性关联较大的正相关峰。结论: 甘草抗氧化活性是多组分联合效应的结果,2、7、10、13、18、21、23号峰为7个与抗氧化活性密切相关的成分,为药材质量控制提供更全面的参考。
Objective: To establish an HPLC fingerprint method of Glycyrrhizae Radix et Rhizoma, and tostudy the relationship between spectrum and antioxidant activity. Methods: Fingerprint by HPLC was established on Glycyrrhizae Radix et Rhizoma from different sources. On the basis that the similaritieswith the reference fingerprintabove 0.915,25 common peaks were evaluated with hierarchical cluster analysis(HCA) and principal component analysis(PCA). The antioxidant activity was determined by 1,1-diphenyl- 2- picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl(DPPH), ferric ion reduction ability plasma(FRAP) analysis, andsalicylic acid assays.Partial least squares regression(PLSR) analysis and grey relational analysis(GRA) were used to study the relationship between HPLC fingerprint and free radical- scavenging activity. Results: The HPLC fingerprint of 25 batches of Glycyrrhizae Radix et Rhizoma was established, and 26 common peaks were determined,with the similarities above 0.915. Nine common peaks were identified by comparing to reference substances.The result of HCA showed that Glycyrrhizae Radix et Rhizoma from different sources were clustered into four categories, which was in consistent with the result of PCA.All of the samples had different degrees of antioxidant activity. The spectrum-activity relationship showed that the peaks 2,7,10,13,18,21 and 23 were positively correlated with the antioxidant activity. Conclusion: The antioxidant activity of Glycyrrhizae Radix et Rhizomais the result of the combined effects of multiple components, and peaks of 7 components are closely related to the antioxidant activity. The present study provides more comprehensive reference for quality control of Glycyrrhizae Radix et Rhizoma.
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