顶羽菊不同极性提取物高效液相指纹图谱与体外抗氧化活性谱效关系研究

Spectrum-effect relationship between HPLC fingerprint and in vitro antioxidant activity of different polar extracts from Rhaponticum repens

关键词:  顶羽菊  不同极性提取物  指纹图谱  抗氧化活性  谱效关系

Key words:  Rhaponticum repens  different polar extract  fingerprint  antioxidant activity  spectrum-effect relationship

摘要:

[目的] 建立顶羽菊不同极性提取物高效液相（HPLC）指纹图谱，并探讨其抗氧化谱效关系，初步明确其抗氧化活性的有效成分。[方法] 采用超高效液相色谱-串联质谱（UPLC-MS/MS）法定性分析顶羽菊化学成分；采用HPLC法建立顶羽菊不同极性提取物的指纹图谱；结合1，1-二苯基-2-苦基肼基（DPPH）自由基清除率和总还原能力的测定方法评价不同提取物的抗氧化活性；利用灰色关联度法和偏最小二乘法联合分析共有峰与抗氧化活性的谱效关系，筛选出具有抗氧化活性的有效成分，并进行单体化合物体外抗氧化实验验证。[结果] 指纹图谱结果显示，顶羽菊8种不同极性提取物HPLC指纹图谱共确定22个共有峰，结合质谱结果指认出其中7个化学成分。抗氧化实验结果显示，不同极性提取物的抗氧化活性存在差异，其中50%乙醇提取物的乙酸乙酯部位和正丁醇部位抗氧化活性最强。谱效关系分析结果显示，1（新绿原酸）、2（绿原酸）、3（隐绿原酸）、5、7、13号共有峰所代表的化学成分对顶羽菊抗氧化活性贡献最大。抗氧化验证结果显示，绿原酸、新绿原酸、隐绿原酸具有抗氧化活性且呈一定浓度依赖性。[结论] 顶羽菊的抗氧化活性是多成分共同起效的结果，有效成分主要为绿原酸、新绿原酸和隐绿原酸，可以为顶羽菊药材的抗氧化药效物质基础、质量控制及开发利用提供参考。

Abstract:

[Objective] To establish the HPLC fingerprints of different polar extracts from Rhaponticum repens and investigate their spectrum-effect relationship with antioxidant activity，preliminarily identifying the effective components contributing to their antioxidant activity. [Methods] Ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）was used to qualitatively analyze the chemical components of Rhaponticum repens. HPLC was used to establish fingerprints of different polar extracts of Rhaponticum repens. The antioxidant activities of different extracts were evaluated by the DPPH radical scavenging rate and total reducing capacity assays. The relationships between common peaks and antioxidant activity were established by partial least squares analysis and gray relational analysis，the effective components of antioxidant activity were screened，and in vitro antioxidant validation tests were carried out. [Results] Twenty-two common peaks were identified from the fingerprint profiles of eight different polar extracts of Rhaponticum repens，and seven chemical components were identified based on mass spectrometry results. The antioxidant activity results showed that different polar extracts exhibited different antioxidant activities. Among them，the ethyl acetate fraction and n-butanol fraction of the 50% ethanol extract had the strongest antioxidant activity. Analysis of the spectrum-effect relationship showed that the chemical components represented by peaks 1（neochlorogenic acid），2（chlorogenic acid），3（cryptochlorogenic acid），5，7，and 13 contributed the most to the antioxidant activity of Rhaponticum repens. The validation results of active ingredients showed that chlorogenic acid，neochlorogenic acid，and cryptochlorogenic acid possessed antioxidant activity in a concentration-dependent manner. [Conclusion] The antioxidant activity of Rhaponticum repens results from the synergistic effects of multiple components，with chlorogenic acid，neochlorogenic acid，and cryptochlorogenic acid being the primary active compounds. These findings provide a reference for elucidating the material basis of its antioxidant efficacy，as well as for quality control and further development and utilization of the medicinal material.