食源性ACE抑制肽体外活性检测方法的改进及应用  

Improvement and application of an analysis method for food-derived ACE inhibitory peptides

作　　者：赵元晖[1] 李八方[1] 刘艺杰 耿娟[3] 曾名勇[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003 [2]青岛出入境检验检疫局,山东青岛266001 [3]山东出入境检验检疫局检验检疫技术中心,山东青岛266002

出　　处：《中国海洋药物》2009年第4期5-8,共4页Chinese Journal of Marine Drugs

基　　金：国家"863"高技术研究发展资助项目(2006AA09Z438)

摘　　要：目的建立一种快速、准确地测定食源性ACE抑制肽体外活性的检测方法。方法通过在不同时间对ACE酶解液中马尿酸含量的测定,确定ACE酶与底物HHL的反应时间;以海参肽为ACE抑制剂,反应液中生成的马尿酸为检测指标,Zorbax SB-C_(18)分析柱为固定相,乙腈/超纯水为流动相,建立一种测定食源性ACE抑制肽体外活性的RP-HPLC法。结果ACE酶与底物HHL的反应时间确定为60min;RP-HPLC的洗脱条件为柱温25℃,流动相乙腈/超纯水的体积比为1:1(各含0.1%三氟乙酸),流速0.4 mL·min^(-1),检测波长228 nm。应用该方法对马尿酸浓度与峰面积的相关性进行分析,结果表明马尿酸浓度在0～200μg·mL^(-1)和200～800μg·mL^(-1)范围内与峰面积呈现良好的相关性;经卡托普利、牡蛎酶解液、鳀鱼酶解液验证,表明该方法不仅适用于食源性ACE抑制肽体外活性的测定,而且也适用于化学合成的降压药物。结论该方法操作简便、重复性好、准确性高,可作为食源性ACE抑制肽体外活性的检测方法。Objective To establish a rapid and accurate analysis method for food-derived ACE inhibitory peptides activity in vitro. Methods Reaction time of ACE and suhstrate was by measuring the hippuric acid liberated in the ACE reaction mixture at regular intervals; An optimal RP-HPLC method to measure food-derived ACE inhibitory peptides activity in vitro was set up. The hippuric acid from ACE reaction mixture （sea cucumber peptides were regarded as ACE inhibitor） was estimated by Zorbax SB-C18 analytical column with acetonitrile and ultrapure water as mobile phase. Results The reaction time of ACE with substrate was determined at sixty minutes; The elution was carried out with the ratio of acetonitrile to ultrapure water was 1 ： 1 （0.1% TFA） at a flow rate of 0.4 mL · min^-1 The absorbance of the eluent was monitored at 228 nm, and column temperature was 25 ℃. The relationship between hippuric acid concentration and peak area exhibited a good linearity in the concentration ranges of 0-200μg· mL^-1 and 200-800 μg· mL^-1. The RP-HPLC method was further validated By captopril, the oyster hydrolysate and the anchovy hydrolysate. Conclusion The method has been proved to be convenient, accurate and suitable for the analysis of foodderived ACE inhibitory peptides activity in vitro.

关 键 词：血管紧张素转化酶 ACE抑制肽 反相高压液相色谱 马尿酸 

分 类 号：R965[医药卫生—药理学]