Development and Validation of HPTLC Method For Simultaneous Determination of Alogliptin and Metformin In Fixed Dose Combination Tablets

A simple, rapid, accurate and precise high performance thin-layer chromatography (HPTLC) method was developed and validated for simultaneous estimation of alogliptin and metformin active pharmaceutical ingredients and fixed dose combination. Alogliptin and metformin densitograms were developed on silica gel 60 F254 HPTLC plates with chloroform: methanol: 0.5 % ammonium sulphate (4:4:2 %v/v) as mobile phase. Densitometric quantification was performed at 254 nm. For Alogliptin and Metformin Rf values were found as 0.66 and 0.44, respectively. The linearity curves of Alogliptin and Metformin were obtained in the concentration range of 100-500 ng/spot and 4000-20000 ng/spot by area with correlation co-efficient of 0.998 and 0.995 for Alogliptin and Metformin, respectively. Limit of detection was found to be 2 ng and 40 ng/spot for Alogliptin and Metformin, respectively; lowest possible quantity to be quantified by the proposed method was found to be 6 ng and 130 ng per spot for Alogliptin and Metformin, respectively. The method was validated for precision, accuracy, specificity and robustness. The developed method was validated and found to be selective, specific and suitable for application in pharmaceutical analysis of these drugs in bulk and fixed dose combination.