SIMULTANEOUS ESTIMATION OF RAMIPRIL AND ITS ACTIVE METABOLITE RAMIPRILAT IN HUMAN PLASMA BY ESI-LC-MS/MS
A rapid and sensitive liquid chromatography tandem mass spectrometry method has been developed and validated for the simultaneous determination of ramipril and ramiprilat in human plasma. The solid-phase extraction technique was used for the extraction of ramipril and ramiprilat from human plasma. Enalapril was used as the internal standard (IS).Chromatography was performed on a Aquasil C18, 100mmÃ—2.1mm, 5Âµ column, with the mobile phase consisting of acetonitrile-deionsed water (in a 65:35 ratio) and 1.0mLL-1 ammonium trifluoroacetate solution (1.0M), followed by detection using mass spectrometry. The method involves a simple reversed isocratic chromatography condition and mass spectrometry detection, which enables detection at sub-nanogram levels. The method was validated and the lower limit of quantification for ramipril and ramiprilat was found to be 0.1 ngmLâˆ’1and 0.1 ngmLâˆ’1, respectively. The mean recovery for ramipril and ramiprilat ranged from 63.5 to 74.3%. This method increased the sensitivity and selectivity; resulting in high-throughput analysis of ramipril and ramiprilat using single IS in a single experiment for bioequivalence studies, with a chromatographic run time of 3.0 min only.
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