A Validated RP-HPLC method for related substances of Dabigatran etexilate mesylate

Abstract

The scope of this research work was to develop a reverse-phase liquid chromatographic method for the quantification of related impurities of dabigatran and to validate the method according to ICH guidelines. Chromatographic conditions were optimised with Poroshell SB C18, 150mm, 4.6mm, 2.7µm particle size column, mixer of Phosphate buffer with phosphoric acid in water and acetonitrile with percentage of 10:90 (v/v) as solvent-A and acetonitrile and buffer with a percentage of 70:30 (v/v) as solvent-B. Gradient compassion mode is employed for mobile phase delivery with a flow rate of 0.8 mL/min. Stationary phase was maintained at 35°C temperature and detection at 230 nm, with 10 µL of sample injection volume. Water and acetonitrile in the percentage 30:70(v/v) were used as a diluent. The developed RP-HPLC method was validated according to ICH guidelines. LOD and LOQ values for dabigatran its impurities were in the range from 33 to 55 ppm and 112 to 168 ppm correspondingly. Method validation results for all the parameters are meeting the ICH guidelines acceptance criteria for the parameters of robustness, ruggedness, linearity, reproducibility and recovery. The proposed method was found to be suitable for the quantitative determination of potential impurities in the bulk samples of Dabigatran etexilate mesylate API.