DEVELOPMENT AND VALIDATION OF LC-MS/MS METHOD FOR THE SIMULTANEOUS DETERMINATION OF PIOGLITAZONE AND VOGLIBOSE IN HUMAN PLASMA

A reliable, simple, robust and sensitive LC-MS/MS (Liquid Chromatography tandem Mass Spectrometric) method has been developed and validated that employs protein precipitation extraction method for the simultaneous determination of Pioglitazone and Voglibose in human plasma by using miglitol as internal standard. Chromatography conditions included an isocratic mobile phase composing of 5 mM ammonium formate buffer: acetonitrile in the ratio 50:50 V/V. the column used was Welchrom XB C18, with specifications of 50×4.6 mm, 5 μm, at a flow rate of 0.80 mL/min. the retention time of Pioglitazone, voglibose and miglitol occurred at ~2.83, 0.8 and 0.70 min, respectively and the total chromatographic run time was 4.0 min. Pioglitazone and voglibose achieved a linear response function in human plasma at 1.07-958 ng/mL & 2.02-203 ng/mL respectively. Pioglitazone and voglibose attained an intra and inter-day accuracy and precision at 0.93 – 7.54% and 0.96 - 5.39%; 0.74 - 7.1 and 0.9 - 3.75% range respectively. the International Conference on Harmonisation of technical Requirements for Registration of Pharmaceuticals for Human Use guidelines were used as the bench mark for validating the method.