Determination of omeprazole in human plasma by high performance chromatography method (HPLC-DAD)

Abstract

     In this study, the HPLC method for determination of omeprazole in human plasma was developed and validated. Plasma samples were cleared of protein by methanol, followed by centrifugation at 12000rpm for 10min. Solutions obtained after centrifugation were filtered through 0.45μm pore size membrane prior to injecting. Chromatography was performed as follows: Column C18 (4.6 x 150mm, 5μm) with corresponding pre-column; Detector - DAD (301nm); Mobile phase: acetonitrile - dipotassium hydrogen phosphate 3mM (27:73; Flow rate 1.2ml/min; The method was validated for repeatability (RSD<5%), accuracy (92.91 - 98.48%), linearity (r>0.999 at 250 - 8000ng/ml) and sensitivity (LOQ = 289.5ng/ml). The high extraction efficiency (94.2 - 98.8%) with RSD<5% was obtained. Plasma samples were stable after 3 cycles of freezing and during sample processing. All proved the method well met the FDA requirements for quantitation of omeprazole in the biological sample. Thus, this analytical procedure can be applied to quantify OMZ in serum human.