A high-performance liquid chromatographic method and a UV derivative spectrophotometric method for the simultaneous determination of metformin (MFN), pioglitazone (PLZ) and glimepiride (GLM), in tablets were developed in the present work. The various parameters, such as linearity, precision, accuracy, specificity, robustness, limit of detection and limit of quantitation were studied according to International Conference on Harmonization guidelines. HPLC was carried out by using the reversed-phase technique on an phenomenex RP-18 column (150x 4.6mm, 5μ) with a mobile phase consisted of acetonitrile and phosphate buffer (pH 3) in the ratio of 65: 35. The flow rate was fixed at 0.5ml/min and the drugs were monitored at 245nm with UV dual absorbance detector and the elution time was found less than 10 min indicates shorter analysis time. The first derivative UV spectrophotometric method was performed at 260.1, 280.7 and 251.5nm for MFN, PLZ and GLM respectively. Statistical analysis was done by Student’s t-test and F-test, which showed no significant difference between the results obtained by the two methods. The proposed methods are highly sensitive, precise and accurate and therefore can be used for its intended purpose.