Abstract
A simple and economical reversed phase HPLC-UV method was developed for omeprazole (OMP) and its two main metabolites, 5-Hydroxy omeprazole (5-OH-OMP) and omeprazole sulfone (OMP-S) in human plasma using pantoprazole as the internal standard. After optimization of various chromatographic conditions and experimental parameters, the method was validated according to standard guidelines. The separation of all the analytes was achieved on a Supelco C18 column using methanol and phosphate buffer (pH 7.2) in the ratio of (42:58, v/v) as the mobile phase in isocratic mode at a flow rate of 0.8 mL/min and wavelength of 302 nm, keeping column oven temperature at 45°C. Extraction of analytes from plasma was achieved through precipitation method followed by injecting a sample volume of 20 µL into the HPLC system. The limits of detection were 3, 1.5, and 1.3 ng/mL and limits of quantification were 10, 5, and 5 ng /mL for 5-0H-OMP, OMP-S, and OMP, respectively. The method was also successfully applied for the determination of S-isomer of omeprazole (esomeprazole) in human plasma.
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