山楂叶提取物抗氧化活性及谱效关系研究  

Relationship Between Antioxidant Activity and Spectrum-Effect of Hawthorn Leaf Extracts

作　　者：李艳荣[1] 周维维 王子怡 赵胜男[1] 潘海峰[1] LI Yan-rong;ZHOU Wei-wei;WANG Zi-yi;ZHAO Sheng-nan;PAN Hai-feng(Hebei Key Laboratory of Study and Exploitation of Chinese Medicine,Chengde Medical College,Chengde 067000,China;Chengde City Food and Drug Inspection and Testing Center,Chengde 067000,China)

机构地区：[1]承德医学院河北省中药研究与开发重点实验室,河北承德067000 [2]承德市食品药品检验检测中心,河北承德067000

出　　处：《中国药学杂志》2020年第20期1673-1679,共7页Chinese Pharmaceutical Journal

基　　金：河北省省级科技计划资助(20372507D);河北省中医药管理局项目资助(2017193);承德医学院自然科学研究计划重点项目资助(201811);河北省高校重点学科建设项目资助(冀教高[2013]4)。

摘　　要：目的比较山楂叶不同提取物的体外抗氧化能力,以抗氧化作用为标准筛选出最优提取物进行总黄酮含量测定和指纹图谱研究,建立谱效关系。方法以维生素C为阳性对照,用1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)清除率来检测不同山楂叶提取物的抗氧化活性,筛选出抗氧化作用最强的提取物。以不同批次筛选出的提取物为研究对象,运用高效液相色谱(UPLC)法,色谱柱为Waters ACQUITY UPLC CSHTM C18(2.1 mm×100 mm,1.7μm),流动相以乙腈-0.1%甲酸水进行梯度洗脱,流速0.15 mL·min-1,柱温30℃,进样量3μL,检测波长320 nm,建立UPLC指纹图谱。采用可见-紫外分光光度法测定总黄酮的含量。用双变量相关性分析、灰色关联法研究其谱效关系。结果4种提取物都有一定抗氧化活性,其中2015年版《中国药典》收录山楂叶提取物清除DPPH和ABTS自由基的能力最强,IC50值与总黄酮的含量有较强的相关性(r=-0.896,r=-0.808),UPLC指纹图谱共标定出33个共有峰,33个共有峰与清除自由基的关联度较高,关联度均大于0.62。结论以"谱-效"结合的思路为山楂叶提取物及其制剂的质量控提供依据。OBJECTIVE To select the optimal extract of Hawthorn leaves based on the antioxidant capacity of different extracts in vitro for the determination of total flavonoids content and the study of fingerprint,and establish the spectrum-effect relationship.METHODS With vitamin C as the positive control,1,1-diphenyl-2-picrylhydrazyl(DPPH)and 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS)clearance rates were used to test the antioxidant activities of different extracts of Hawthorn leaves,and the extract with the strongest antioxidant activity was selected and used as the research object.UPLC analysis for determination of the content of total flavonoids was performed on Waters ACQUITY UPLC CSHTM C18(2.1 mm×100 mm,1.7μm)column using 0.1%formic acid water and acetonitrile as mobile phase.Gadient elution was conducted at the flow rate of 0.15 mL·min-1,the injection volume was 3μL,the column temperature was 30℃,and the detection wavelength was 320 nm.The UPLC fingerprint was established.The spectrum-effect relationship was studied by using bivariate correlation analysis and grey correlation method.RESULTS Four kinds of extracts had certain antioxidant activity,among which hawthorn leaf extract listed in the Chinese Pharmacopoeia 2015 edition had the strongest ability to scavenge DPPH and ABTS free radicals(r=-0.896,r=-0.808).A total of 33 peaks were identified by UPLC fingerprint.They were highly correlated with the scavenging ability for free radicals and the correlation coefficients were all greater than 0.62.CONCLUSION This study provided the basis for the quality control of Hawthorn leaf extract and its preparations.

关 键 词：山楂叶提取物 抗氧化活性 总黄酮 指纹图谱 谱效关系 

分 类 号：R284[医药卫生—中药学]