Journal
EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES
Volume 101, Issue -, Pages 182-188Publisher
ELSEVIER
DOI: 10.1016/j.ejps.2017.02.015
Keywords
Angiotensin-converting enzyme; Angiotensin I; Captopril; Lisinopril; Inhibition; LC-QqQ-MS; Multiple reaction monitoring (MRM)
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Funding
- Higher Education Commission (HEC), Pakistan
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Angiotensin-converting enzyme (ACE) plays a key role in regulating blood pressure in the body by converting the angiotensin I (AI) into angiotensin II (AII). Angiotensin II is a potent vaso-active peptide that causes arterioles to constrict, resulting in increased blood pressure. A rapid and sensitive method for the identification of inhibitors of ACE was developed, and optimized employing HPLC-ESI-QqQ-MS. In this assay, angiotensin I substrate was converted into the product angiotensin II with the catalytic action of ACE. A calibration curve for depleting concentration of angiotensin I was developed and linearity of R-2 = 0.999 with a remarkably low concentration of substrate range 20-200 nM. The limit of detection and quantification of angiotensin I was found to be 1.93 and 5.84 nM, respectively. The enzymatic reaction was optimized for incubation time, concentration, and volume of enzyme and substrate. All reactions were performed at 37 degrees C at pH 7.5 with standard incubation time of 20 min. Two standard inhibitors, Captopril and Lisinopril, were checked through the newly developed method for their inhibitory potential, and their IC50 values were found to be 3.969 and 0.852 mu M, respectively. Reproducibility and precision analysis of different experiments, showed < 9.9% RSD. The developed method can be,used for the identification of new ACE inhibitors. (C) 2017 Elsevier B.V. All rights reserved.
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