黄连花薹HPLC指纹图谱的建立及其抗氧化和抑菌作用谱效关系研究
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篇名: 黄连花薹HPLC指纹图谱的建立及其抗氧化和抑菌作用谱效关系研究
TITLE: Establishment of HPLC Fingerprint of Coptis chinensis Inflorescence and Its Spectrum-effect Relationship with Antioxidant and Antibacterial Effects
摘要: 目的:建立黄连花薹的高效液相色谱(HPLC)指纹图谱,并研究其与抗氧化和抑菌作用的谱效关系。方法:以14批不同产地的黄连花薹为对象,采用HPLC法测定。色谱柱为SupersilC18,流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为1.0mL/min,柱温为25℃,检测波长为329nm,进样量为10μL。采用《中药指纹图谱相似度评价系统》(2012A版)建立14批黄连花薹的指纹图谱并进行相似度评价和共有峰指认。以DPPH自由基清除率和羟自由基清除率为抗氧化作用指标,以相对抑菌活性(大肠埃希菌)为抑菌作用指标,利用SPSS21.0软件对黄连花薹共有峰与上述两种药效指标进行Pearson相关性分析,建立谱效关系模型,并进行验证。结果:黄连花薹的HPLC指纹图谱中共得到7个共有峰,相似度均不低于0.916,并指认5号峰为盐酸小檗碱。7个共有峰均与DPPH自由基清除率呈正相关;1~3、4、6、7号峰与羟自由基清除率呈正相关,5号峰与羟自由基清除率呈负相关;5号峰与相对抑菌活性呈正相关。经验证,黄连花薹样品的DPPH自由基清除率、羟自由基清除率、相对抑菌活性的预测值与测量值的相对误差为0.92%~14.5%。结论:所建立的谱效关系模型可用于黄连花薹抗氧化、抑菌作用的评价;1、2、3、4、6、7号峰所代表化学成分是黄连花薹抗氧化作用的物质基础,盐酸小檗碱是其抑菌作用的物质基础。
ABSTRACT: OBJECTIVE:To establish HPLC fingerprint of Coptis chinensis inflorescence,and study its spectrum-effect relationship with antioxidant and antibacterial effects. METHODS :Taking 14 batches of C. chinensis inflorescence from different producing areas as the object ,HPLC method was adopted. The determination was performed on Supersil C 18 column with mobile phase consisted of acetonitrile- 0.1% phosphoric acid solution(gradient elution )at the flow rate of 1.0 mL/min. The column temperature was set at 25 ℃. The detection wavelength was set at 329 nm,and sample size was 10 μL. The fingerprints of 14 batches of C. chinensis inflorescence were established by Similarity Evaluation System of TCM Fingerprint (2012 A edition ),and the similarity evaluation and common peak identification were carried out. Taking DPPH free radical scavenging rate and hydroxyl radical scavenging rate as antioxidant effects index ,relative antibacterial activity (Escherichia coli )as antibacterial effect index , SPSS 21.0 software was adopted to analyze the Pearson correlation between common peaks of C. chinensis inflorescence and above efficacy indexes ;their spectrum-effect relationship was established and validated. RESULTS :A total of 7 common peaks were obtained in HPLC fingerprint of C. chinensis inflorescence,and the similarity was no less than 0.916. No. 5 peak was identified as berberine hydrochloride. Seven common peaks were positively correlated with DPPH free radical scavenging rate ;No. 1-3,4,6,7 peaks were positively correlated with hydroxyl radical scavenging rate ,while No. 5 peak was negatively correlated with hydroxyl radical scavenging rate. There was a positive correlation between No. 5 peak and antibacterial activity in vitro . After validation , relative error between the predicted values and the measured values of DPPH free radical scavenging rate ,hydroxyl radical scavenging rate and relative antibacterial activity was 0.92%- 14.5% . CONLUSIONS :The established spectrum-effect relationship model can be used to evaluate antioxidant andantibacterial effects of C. chinensis inflorescence. The chemical components represented by No. 1,2,3,4,6,7 peaks are the material basis of antioxidant effect of C. chinensis inflorescence, and berberine hydrochloride is the material basis of antibacterial effect.
期刊: 2021年第32卷第05期
作者: 杨慧,张佳,刘博文,刘钱,李松洋,吴磊,徐玉玲,刘涛
AUTHORS: YANG Hui,ZHANG Jia,LIU Bowen ,LIU Qian,LI Songyang ,WU Lei,XU Yuling ,LIU Tao
关键字: 黄连花薹;高效液相色谱;指纹图谱;抗氧化作用;抑菌作用;物质基础
KEYWORDS: Coptis chinensis inflorescence ;HPLC;Fingerprint;Antioxidant activity ;Antibacterial activity ;Material basis
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