Page 16 - 2020年1月第31卷第2期

啤酒花的HPLC指纹图谱建立及其抗氧化作用谱效关系研究 Δ

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刘晓燕 ，蒋益萍 ，张嘉宝 ，王娜妮 ，辛海量 （1.海军军医大学药学院，上海 200433；2.浙江省中医药研究
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院，杭州 310007）
中图分类号 R931 文献标志码 A 文章编号 1001-0408（2020）02-0138-06
DOI 10.6039/j.issn.1001-0408.2020.02.03

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Diamonsil C18，
流动相为0.1％磷酸水溶液-乙腈（梯度洗脱），流速为1.0 mL/min，柱温为30 ℃，检测波长为358 nm，进样量为2 μL。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SIMCA 14.1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，0.1 mg/mL黄腐酚与0.01 mg/mL
维生素C对DPPH自由基的清除作用相当，但10.0 mg/mL黄腐酚对ABTS自由基清除作用小于0.1 mg/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。
关键词 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

Establishment of HPLC Fingerprint and Its Spectrum-effect Relationship Study of Humulus lupulus
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LIU Xiaoyan ，JIANG Yiping ，ZHANG Jiabao ，WANG Nani ，XIN Hailiang（1. College of Pharmacy，Naval
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Medical University，Shanghai 200433，China；2. Zhejiang Institute of Traditional Chinese Medicine，Hangzhou
310007，China）
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 C18 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
KEYWORDS Humulus lupulus；HPLC；Fingerprint；Antioxidant；Spectrum-effect relationship；PLSR

Δ 基金项目：国家自然科学基金资助项目（No.U1603283） 啤酒花（Humulus lupulus L.）为桑科葎草属多年生
＊硕士研究生。研究方向：中药药理学。E-mail：nmulxy@163. 草质蔓生藤本植物，主要分布于我国新疆地区，以未成
com
熟的雌性球穗花序入药，具有健胃消食、安神、利尿等功
# 通信作者：副教授，硕士生导师，博士。研究方向：中药资源、中 [1]
药（抗骨质疏松）药理学。电话：021-81871300。E-mail：hailiangxin@ 效，可用于治疗肺结核、癔病、失眠、膀胱炎等症 ，在我
163.com 国作为民族药收载于1977年版《中国药典》（一部）、《新

·138 · China Pharmacy 2020 Vol. 31 No. 2 中国药房 2020年第31卷第2期