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

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

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.

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.