Liquid Chromatographic Assay of Peptides Activity with Inhibiting Angiotensin Converting Enzyme

高等学校化学研究 ›› 2010, Vol. 26 ›› Issue (5): 712-716.

Liquid Chromatographic Assay of Peptides Activity with Inhibiting Angiotensin Converting Enzyme

LIU Jing-bo1, YU Zhi-peng1, ZHAO Wen-zhu1, LU Jing1, CHEN Feng2 and LIN Song-yi1*

1. Laboratory of Nutrition and Functional Food, Jilin University, Changchun 130062, P. R. China;
2. Department of Food Science and Human Nutrition, Clemson University, Clemson SC 29634, USA

收稿日期:2010-03-01 修回日期:2010-07-06 出版日期:2010-09-25 发布日期:2010-11-26
通讯作者: LIN Song-yi. E-mail: linsongyi730@163.com
基金资助:
Supported by the National High Technology Research and Development Program of China(No.2007AA10Z329).

Liquid Chromatographic Assay of Peptides Activity with Inhibiting Angiotensin Converting Enzyme

LIU Jing-bo1, YU Zhi-peng1, ZHAO Wen-zhu1, LU Jing1, CHEN Feng2 and LIN Song-yi1*

1. Laboratory of Nutrition and Functional Food, Jilin University, Changchun 130062, P. R. China;
2. Department of Food Science and Human Nutrition, Clemson University, Clemson SC 29634, USA

Received:2010-03-01 Revised:2010-07-06 Online:2010-09-25 Published:2010-11-26
Contact: LIN Song-yi. E-mail: linsongyi730@163.com
Supported by:
Supported by the National High Technology Research and Development Program of China(No.2007AA10Z329).

摘要/Abstract

摘要： A rapid, simple and interference-free method was developed to evaluate the inhibitory activity of hydrolyzed peptides from egg white protein against the angiotensin-converting enzyme. The total reaction volume was 60 μL, saving the cost. The assay was based on a HPLC separation and quantification of the synthetic substrate hippuryl-L-histidyl-L-leucine and its hydrolyzed product―hippuric acid; the separation was performed on a C18 column eluted by a mobile phase of acetonitrile/water(0.5% TFA) at a volume ratio of 25:75. At a signal to noise ratio(S/N) of 10, the detective limit of the quantitation of hippuric acid was (0.4600±0.0097) μmol/L. The standard curve shows a linear response with a slope of 49488 and a correlation coefficient of 0.9995. The assay was adequate for the study of ACE inhibition by Captopril and peptides derived from food protein, and showed a very good correlation with the previous methods.

关键词: Angiotensin converting enzyme, HPLC, Egg white protein, Peptide

Abstract: A rapid, simple and interference-free method was developed to evaluate the inhibitory activity of hydrolyzed peptides from egg white protein against the angiotensin-converting enzyme. The total reaction volume was 60 μL, saving the cost. The assay was based on a HPLC separation and quantification of the synthetic substrate hippuryl-L-histidyl-L-leucine and its hydrolyzed product―hippuric acid; the separation was performed on a C18 column eluted by a mobile phase of acetonitrile/water(0.5% TFA) at a volume ratio of 25:75. At a signal to noise ratio(S/N) of 10, the detective limit of the quantitation of hippuric acid was (0.4600±0.0097) μmol/L. The standard curve shows a linear response with a slope of 49488 and a correlation coefficient of 0.9995. The assay was adequate for the study of ACE inhibition by Captopril and peptides derived from food protein, and showed a very good correlation with the previous methods.

Key words: Angiotensin converting enzyme, HPLC, Egg white protein, Peptide

LIU Jing-bo1, YU Zhi-peng1, ZHAO Wen-zhu1, LU Jing1, CHEN Feng2 and LIN Song-yi1. Liquid Chromatographic Assay of Peptides Activity with Inhibiting Angiotensin Converting Enzyme[J]. 高等学校化学研究, 2010, 26(5): 712-716.

LIU Jing-bo1, YU Zhi-peng1, ZHAO Wen-zhu1, LU Jing1, CHEN Feng2 and LIN Song-yi1*. Liquid Chromatographic Assay of Peptides Activity with Inhibiting Angiotensin Converting Enzyme[J]. Chemical Research in Chinese Universities, 2010, 26(5): 712-716.

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Liquid Chromatographic Assay of Peptides Activity with Inhibiting Angiotensin Converting Enzyme

Liquid Chromatographic Assay of Peptides Activity with Inhibiting Angiotensin Converting Enzyme

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