Development and Validation of a Method for Simultaneous Determination of Metformin Hydrochloride and Sitagliptin Phosphate in a Formulation by RP-HPLC

P. B. N. Prasad, K. Satyanarayana, G. Krishnamohan

Central Drugs Testing Laboratory, Hyderabad, India.
Department of Pharmacy, JNTUH, Hyderabad, India.
NATCO Pharma R&D, Hyderabad, India.

DOI: 10.4236/ajac.2014.511082                

Abstract

Present study was aimed to develop and validate a reverse-phase high-performance liquid chromatography method for simultaneous determination of sitagliptin phosphate and metformin hy-drochloride in a marketed formulation. The drug separation was performed on Hibar-240, Li-chrosphere-100 C18 ODS (250 × 4.6 mm, 5 μm) column, at a flow rate of 1 mL/min. The mobile phase used was a mixture of methanol: potassium di-hydrogen phosphate buffer at a ratio of 70:30 v/v. The detection was carried out at a wavelength of 266 nm. The retention times of sitagliptin phosphate and metformin hydrochloride were found as 6.1 and 4.9 min respectively. Linear calibration curves with good correlation coefficients were obtained over the concentration ranges of 10 - 50 μg/mL for sitagliptin and 20 - 100 μg/mL for metformin. The limit of detection was 0.016 and 0.14 μg/mL and the limit of quantification was 0.048 and 0.42 μg/mL for sitagliptin phosphate and metformin hydrochloride respectively. Validation of the method demonstrated system selectivity, specificity, linearity, accuracy and precision. The developed method was found useful in the simultaneous analysis of sitagliptin phosphate and metformin hydrochloride in formulation.

Keywords

Sitagliptin Phosphate, Metformin Hydrochloride, Reverse Phase High Performance Liquid Chromatography (RPHPLC), International Committee on Harmonization (ICH) Guidelines, Limit of Detection (LOD), Limit of Quantitation (LOQ), Linearity, Precision, Accuracy & Reproducibility

Conflicts of Interest

The authors declare no conflicts of interest.

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