A simple, rapid, sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for quantification of Ramipril and its metabolite Ramiprilat in K2 EDTA human plasma. Both the drugs were extracted by solid phase extraction. Enalpril was used as the internal standard (IS). The chromatographic separation was performed by binary gradient on HYPURITY C18, 3x50 mm,3µ with a mobile phase of 0.1% formic acid solution in water: organic mixture (10:90) (v/v)[organic mixture-methanol:acetonitrile(90:10)] followed by detection using mass spectrometry. The protonated analyte was quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The method was validated and the lower limit of quantification for Ramipril and Ramiprilat was found to be 0.390 ngmL−1and 0.395 ngmL−1 respectively. The mean recovery for Ramipril and Ramiprilat ranged from 95.63 to 85.66% and also this method no significant matrix effect on analytes. The method was validated over the concentration range of 0.300 ng/mL to 40.000 ng/mL for both Ramipril and Ramiprilat in human plasma with a accuracy of within run and between run in the range of 80-120%. The within run and between run precision of Ramipril and Ramiprilat were also within the range of 20% (for LLOQ level) and 15% (for other than LLOQ) of % CV.
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