Reverse Phase Chromatographic Method of Analysis for Assay and Content Uniformity Estimation of Drug Substance Sitagliptin, Metformin and Empagliflozin from Available Marketed Formulation

Abstract

A new method of analysis with reverse phase chemistry was designed and developed. Validation for method of analysis was performed for its intended use to calculate assay and content uniformity of drug substance sitagliptin, metformin and empagliflozin in the drug products. The method has a run time of 10 minutes on X-bridge C18 column having 250 mm length, 4.6 mm internal diameter and Particle Size of 5µm, by the use of 0.1% Trifluoroacetic acid Buffer 40%: Methanol 40%: Acetonitrile 20% ratio as constituent composition in the proposed mobile phase and chromatography run at wavelength of 224 nm. The retention time of Metformin, Empagliflozin and Sitagliptin, were 3.383, 5.571 and 6.429 minutes, respectively. International Conference on Harmonization guideline was referred for validation. The method showed adequate sensitivity for precision, linearity and accuracy parameter (between the range 25-75μg/mL, 250-750μg/mL and 2.5-7.5μg/mL for sitagliptin, metformin and empagliflozin respectively). The percentage recoveries obtained for sitagliptin, metformin and empagliflozin are in the range of 98.0 – 102.0 %. As results are within the acceptance [1], hence the new developed and proposed method is suitable for quantification of one, two or three component drugs, separately or in combination.