DETERMINATION AND VALIDATION OF A HPLC METHOD WITH UV DETECTIONOF ITOPRIDE HYDROCHLORIDE IN HUMAN SERUM
Journal Title: International Journal of Pharmacy and Pharmaceutical Sciences - Year 2015, Vol 7, Issue 11
Abstract
Objective: Determination and validation of a simple, sensitive and rapid reversed phase high-performance liquid chromatography method for the determination of itopride hydrochloride in human serum using nimesulide as an internal standard.Methods: A liquid-liquid extraction method was employed to extract itopride hydrochloride from serum matrix with diethyl ether. Chromatographic separation was obtained within 10 min using a reverse phase C18 column (250 mm×4.6 mm, 5 µm) with an isocratic elution. The mobile phase consisting of a mixture acetonitrile and 0.05M phosphate buffer with a flow rate of 1.0 ml/min was used and the eluents were monitored at 258 nm.Results: Calibration curves were linear (r2= 0.9987) in the range of 10ng/ml to 1000 ng/ml. The limit of quantitation was 14ng/ml. The mean recovery of itopride hydrochloride and the nimesulide from the biological matrix was more than 66.04 and 64.57%, respectively. The inter-day accuracy of the drug containing serum samples was more than 97.81% with a precision of 2.31–3.68%. The intra-day accuracy was 96.91% or more with a precision of 5.17–9.50%.Conclusion: The developed method was validated and successfully applied to a bioequivalence study in healthy human volunteers.Â
Authors and Affiliations
Sreenivas Patro Sisinthy, Selvakumar Duraipandi, Nalamolu Koteswara Rao, Me Bhanoji Rao
Keywords
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