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Analysis method of the angiotensin-I converting enzyme inhibitory activity based on micellar electrokinetic chromatography.
Authors:Toshiro Watanabe  Tapan Kumar Mazumder  Shiro Nagai  Keisuke Tsuji  Shigeru Terabe
Institution:Yaegaki Technology Development Laboratories, Yaegaki Bio-industry, Inc., 681 Hayashida, Himeji, Hyogo 679-4298, Japan. watnb-ts@yaegaki.co.jp
Abstract:A micellar electrokinetic chromatography (MEKC) method was developed for estimating the angiotensin-I converting enzyme (ACE) inhibitory activity by separating the hippuric acid liberated in the ACE reaction mixture in the presence of an inhibitor, captopril. The hippuric acid was successfully separated and detected by MEKC with a 25 mM sodium dodecyl sulfate solution in a 25 mM phosphate-50 mM borate buffer at pH 7.0; the total analysis took about 5 min. A good linear relationship was observed between the inhibitor and the peak area of hippuric acid release. No significant difference in the ACE inhibitory activity (IC50) of captopril (an antihypertensive medicine) or autolyzed-mushrooms (functional foods) was observed between the conventional method and the MEKC method. The MEKC method was found to be a useful technique for a rapid assay of the ACE inhibitory activity.
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