高效液相色谱法测定霉茶成分及其对血管紧张肽转化酶的抑制作用  

Determination of Components from Ampelopsis Grossedentata by HPLC and Inhibitory Effect of Its on Angiotensin Converting Enzyme

作　　者：吴勇[1,2] 罗弘[1] 王瑞[1,2] 谭永霞[1,2] 胡俊杰[1,2] 张宝徽[1,2] 田先翔[1,2] 郑国华[1,2] 张筱祎 

机构地区：[1]湖北中医药大学药学院,武汉430065 [2]中药资源与中药复方省部共建教育部重点实验室,武汉430065

出　　处：《医药导报》2015年第12期1559-1562,共4页Herald of Medicine

基　　金：国家自然科学基金资助项目(81173502)

摘　　要：目的利用高效液相色谱法测定霉茶成分对血管紧张肽转化酶(ACE)的抑制作用。方法以马尿酰-组氨酰-亮氨酸(HHL)为反应底物,经ACE催化生成的马尿酸为测定指标,分别考察霉茶总黄酮、二氢杨梅素、杨梅素对ACE的抑制作用。采用Welchrom C18色谱柱(4.6 mm×250 mm,5μm),流动相为水(含0.1%甲酸,0.1%三乙胺)∶乙腈(含0.1%甲酸,0.1%三乙胺)=78∶22,柱温30℃,流速1.0 m L·min^(-1),检测波长228 nm,进样量10μL。结果马尿酸在0.05~5.00μg范围内其进样量与峰面积呈良好的线性关系(r=0.999 9),霉茶总黄酮、二氢杨梅素对ACE抑制率>75%。结论经该法检测,霉茶总黄酮、二氢杨梅素对ACE均有较强抑制作用。该方法简单、灵敏、重复性好,可用于ACE抑制药的体外活性筛选。Objective To measure the inhibition of components from Ampelopsis grossedentata on angiotensin converting enzyme（ ACE） by high performance liquid chromatograph. Methods By quantitative determination of hippuric acid,the ACE-catalyzed product from the substrate HIPPURYL-HIS-LEU（ HHL）, the inhibitory effects of the total flavonoids of Ampelopsis grossedentata,dihydromyricetin and myricetin were evaluated respectively. High performance liquid chromatograph analysis was conducted on a Welchrom C18column（ 4.6 mm×250 mm,5 μm）,eluting with water ： acetonitrile（ each containing0.1% formic acid and 0.1% triethylamine） = 78∶ 22,at flow rate of 1.0 m L·min-1 under temperature of 30 ℃,the detection wavelength was set at 228 nm,and the injection volume was 10 μL. Results The injection amount of hippuric acid showed good linearity with peak area within the range of 0.05 – 5.00 μg（ r = 0.999 9）,and the inhibitory rate of the total flavonoids of Ampelopsis grossedentata and dihydromyricetin reached more than 75% against ACE. Conclusion As the method exhibited,the total flavonoids of Ampelopsis grossedentata and dihydromyricetin have potent inhibitions against ACE. For being simple,sensitive and repeatable,this method can be used for the screening of ACE inhibitors in vitro.

关 键 词：霉茶成分 血管紧张肽转化酶 抑制作用 含量测定 色谱法 高效液相 

分 类 号：R972.4[医药卫生—药品] R965[医药卫生—药学]