Development And Validation Of RP-HPLC Method For Anagliptin And Metformin Hydrochloride And Its Related Impurities In Tablet Dosage Form

Abstract

Introduction: Anagliptin (ANA) and metformin HCl (MET) are key antidiabetic medications for managing type 2 diabetes by regulating blood glucose through different mechanisms. Despite various analytical methods, a stability-indicating RP-HPLC method for quantifying both drugs and their related impurities is lacking. This study develops and validates a precise, sensitive RP-HPLC method for determining anagliptin, metformin, and anagliptin impurity A, as accordance Q2 (R1) guidelines.

Objectives:Reverse-phase high-performance liquid chromatography (RP-HPLC) method has been developed and validated for estimating the impurities of metformin HCl and anagliptin in a combined tablet dosage form.

Methods:The RP-HPLC analysis utilized a KROMASIL-C₁₈ column (250mm x 4.6mm, 5µm particle size), with a mobile phase of acetonitrile and 0.05M potassium dihydrogen phosphate buffer (pH 3.0) in a 50:50 ratio. The detection was carried out at 220 nm with a flow rate of 1.0 mL/min.This analytical method was validated according to the International Council for Harmonization (ICH) guidelines.

Results: The linearity of metformin HCl (25-75µg/ml) with LOQ 2.99µg/ml, anagliptin (5-15µg/ml) with LOQ 1.304µg/ml andanagliptin Impurity A (5-15µg/ml) with LOQ 1.063µg/ml and Limit of Detection was found to be 0.43µg/ml for anagliptin,0.97µg/ml for Metformin HCl and 0.35µg/ml for impurity A. The correlation coefficient was more than 0.999 for Metformin HCl, anagliptin and Impurity A. The relative standard deviation value for repeatability, Interday precision and intraday precision was less than 2% which indicate the method is robust in nature.Conclusion:In summary, this RP-HPLC method is linear, sensitive, precise, accurate, and robust, making it highly suitable for the quantitative determination of metformin HCl, anagliptin, in presence of anagliptin impurity A in combined tablet dosage form.