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Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Enalapril and Enalaprilat in Human Plasma

Srinivasa Rao and Gopinath Chakka

A fast, perceptive, and highly discriminating Liquid chromatographic method with Mass spectroscopy was urbanized and validated for simultaneous determination of Enalapril and its major bioactive metabolite Enalaprilat in human plasma. Solid phase extraction process was used for the extraction of analytes from plasma. The chromatographic severance was achieved on Zorbax Eclipse; 150 × 4.6 mm, C18 5 µm column using a solvent system of acetonitrile and 0.1% v/v HCOOH in water with a ratio of 65:35 v/v at a flow rate of 0.8 mL/min. The analytes were detected in a positive ionization by multiple reactions monitoring mode. Mass transitions of m/z 377.10 → 234 for Enalapril, m/z 382.10 → 239.20 for Enalapril D5 and m/z 349 → 206 for Enalaprilat, m/z 354.20 → 211.20 for Enalaprilat D5 were used for quantification in plasma samples. The method exhibited a linear response with a Correlation co-efficient (r2) of >0.99 in the concentration range of 0.502-160.2 ng/mL for Enalapril and 0.506-161.5 ng/mL for Enalaprilat in human plasma. The mean recovery of Enalapril was 91.21% with a precision range of 1.72% to 5.06% and Enalaprilat was 90.85% with a precision range of 1.29% to 3.87%. The proposed method can be utilized for the quantification of Enalapril and Enalaprilat in human plasma in regular bioequivalence studies.