高等学校化学学报 ›› 2012, Vol. 33 ›› Issue (11): 2417-2421.doi: 10.7503/cjcu20120151

• 分析化学 • 上一篇    下一篇

溶剂诱导相变萃取-分光光度法测定血管紧张素转化酶活性

田清青1,2, 曾伟秀1, 赵昕1, 侯玉兰1, 陈波1, 姚守拙1   

  1. 1. 湖南师范大学化学生物学中药分析省部共建教育部重点实验室, 长沙 410081;
    2. 湖南中医药高等专科学校, 株洲 412008
  • 收稿日期:2012-02-23 出版日期:2012-11-10 发布日期:2012-10-15
  • 通讯作者: 陈波,男,博士,教授,博士生导师,主要从事中药及色谱分析研究.E-mail:dr-chenpo@vip.sina.com E-mail:dr-chenpo@vip.sina.com
  • 基金资助:

    国家自然科学基金(批准号: 20927005, 21275049, 81274178);国家"八六三"计划项目(批准号: 2010AA023001); 湖南省科研条件创新专项重点项目(批准号: 2010TT1001)和湖南省高校科技创新团队支持计划资助.

Solvent Induced Phase Separation Extraction-Spectrophotometric Method for Determination of Angiotensin-converting Enzyme Activity

TIAN Qing-Qing1,2, ZENG Wei-Xiu1, ZHAO Xin1, HOU Yu-Lan1, CHEN Bo1, YAO Shou-Zhuo1   

  1. 1. Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research, Ministry of Education, Hunan Normal University, Changsha 410081, China;
    2. Hunan Traditional Chinese Medical college, Zhuzhou, 412008, China
  • Received:2012-02-23 Online:2012-11-10 Published:2012-10-15

摘要:

在传统体外测定血管紧张素转化酶(ACE)活性方法的基础上, 以猪肺ACE粗提物为酶源, 采用溶剂诱导相变萃取法(SIPSE)萃取酶反应产物马尿酸(HA), 用分光光度法快速定量测定HA以评估ACE活性或样品对ACE的抑制活性. 以卡托普利为模型验证了此方法, 并分别测定了中药复方天麻钩藤饮及其含有的11味中药对ACE的抑制活性. 结果表明, 猪肺ACE粗提物即可满足ACE抑制活性测定的需要; 与传统液液萃取法相比, SIPSE对HA具有更高的选择性及萃取能力, 萃取率为(97.47±0.55)%(n=3); 通过计算校正, 溶剂诱导相变萃取-分光光度法能简便、 快速且准确地测定成分复杂体系的ACE抑制活性.

关键词: 溶剂诱导相变萃取, 血管紧张素转化酶, 分光光度法, 活性

Abstract:

On the basis of the classical method of the determination of angiotensin-converting enzyme (ACE) activity, the solvent induced phase separation extraction (SIPSE) method and spectrophotometry were employed for the extraction and determination of hippuric acid (HA) formed during enzymatic reaction. Captopril was used as the model for the verification of the developed method. Based on the method, a traditional chinese prescription (Gastrodia and Uncaria Decoction) and the 11 kinds of main medicinal ingredients in the prescription were measured for ACE inhibition. The results show that the SIPSE method is highly selective and efficient in extracting of HA, and the recovery of HA is (97.47±0.55)%(n=3). By necessary correction, the SIPSE-spectrophotometric method is accurate and sufficient for the direct evaluation of the ACE inhibitory activity of high-matrix samples.

Key words: Solvent induced phase separation extraction(SIPSE), Angiotensin-converting enzyme, Spectrophotometry, Activity

中图分类号: