啤酒花的HPLC指纹图谱建立及其抗氧化作用谱效关系研究
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篇名: 啤酒花的HPLC指纹图谱建立及其抗氧化作用谱效关系研究
TITLE: Establishment of HPLC Fingerprint and Its Spectrum-effect Relationship Study of Humulus lupulus
摘要: 目的:建立啤酒花的高效液相色谱(HPLC)指纹图谱,并考察与其抗氧化作用的相关性。方法:色谱柱为DiamonsilC18,流动相为0.1%磷酸水溶液-乙腈(梯度洗脱),流速为1.0mL/min,柱温为30℃,检测波长为358nm,进样量为2μL。以黄腐酚峰为参照,绘制11批啤酒花药材样品的HPLC指纹图谱;采用《中药色谱指纹图谱相似度评价系统》(2012版)进行相似度评价,确定共有峰;以1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率、2,2′-连氨(-3-乙基苯并噻唑啉-6-磺酸)二氨盐(ABTS)自由基清除率为抗氧化作用的药效指标,采用SIMCA14.1软件进行偏最小二乘回归分析以建立谱效关系,并进行体外抗氧化试验验证。结果:11批啤酒花药材样品共有19个共有峰,相似度均大于0.830。共指认出7个成分,分别为黄腐酚、类葎草酮、葎草酮、加葎草酮、类蛇麻酮、蛇麻酮和加蛇麻酮。11批啤酒花均具有清除DPPH和ABTS自由基的作用。谱效关系分析结果显示,啤酒花中黄腐酚、葎草酮、类蛇麻酮峰与其抗氧化能力呈正相关,且变量投影值均大于1。体外抗氧化验证结果显示,0.1mg/mL黄腐酚与0.01mg/mL维生素C对DPPH自由基的清除作用相当,但10.0mg/mL黄腐酚对ABTS自由基清除作用小于0.1mg/mL维生素C。结论:所建HPLC指纹图谱可用于评价啤酒花的质量;黄腐酚、葎草酮和类蛇麻酮可能是啤酒花具有抗氧化作用的物质基础。
ABSTRACT: OBJECTIVE:To establish HPLC fingerprint of Humulus lupulus ,and to investigate its correlation with the antioxidant activity. METHODS :The determination was performed on Diamonsil C 18 column with mobile phase consisted of 0.1% phosphoric acid solution-acetonitrile (gradient elution )at the flow rate of 1.0 mL/min. The column temperature was 30 ℃,and detection wavelength was set at 358 nm,with sample size of 2 μL. Using xanthohumol as reference,HPLC fingerprints of 11 batches of H. lupulus were determined. The similarity of 11 batches of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System (2012 edition)to confirm common peaks. Using scavenging rate of DPPH and ABTS free radical as pharmacodynamic indicators of antioxidant effects ,SIMCA 14.1 analysis software was used for PLSR to establish the spectra-effect relationship ,and validation test of in vitro anti-oxidation was carried out. RESULTS :There were 19 common peaks in HPLC fingerprints of 11 batches of sample ,the similarity of which was higher than 0.830. Seven components were identified as xanthohumol,cohumulone,humulone,adhumulone,colupulone,lupulus and adlupulus. Eleven batches of H. lupulus had the ability to scavenge DPPH and ABTS free radicals. The spectrum-effect relationship showed that xanthohumol ,humulone and colupulone peaks were positively associated with its anti-oxidant ability ,and variable projection value was greater than 1. In vitro antioxidant results showed that scavenging effect of 0.1 mg/mL xanthohumol to DPPH free radicals was similar to that of 0.01 mg/mL vitamine C ,but scavenging effect of 10.0 mg/mL xanthohumol to ABTS free radical was less than that of 0.1 mg/mL vitamin C. CONCLUSIONS:Established HPLC fingerprint can be used for the quality evaluation of H. lupulus ;xanthohumol,humulone and colupulone are the main material basis for the antioxidant effect of H. lupulus
期刊: 2020年第31卷第02期
作者: 刘晓燕,蒋益萍,张嘉宝,王娜妮,辛海量
AUTHORS: LIU Xiaoyan ,JIANG Yiping ,ZHANG Jiabao,WANG Nani,XIN Hailiang
关键字: 啤酒花;高效液相色谱法;指纹图谱;抗氧化;谱效关系;偏最小二乘法回归分析
KEYWORDS: Humulus lupulus ;HPLC;Fingerprint;Antioxidant;Spectrum-effect relationship ;PLSR
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