Method Validation for Stability Indicating Method of Related Substance in Active Pharmaceutical Ingredients Dabigatran Etexilate Mesylate by Reverse Phase Chromatography

Abstract

To develop the stability indicating, highly accurate precise and linear method for related substance of the Dabigatran through the reverse phase high performance liquid chromatography and it is validated as per the current ICH guideline. The optimize method uses a reverse phase column, Poroshell 120 EC-18 (150 mm × 4.6 mm, 2.7µ), Mobile Phase of hexane-1 Sulfonic acid sodium salt monohydrate (6.5 ± 0.05) and Methanol through gradient flow rate of 0.6ml/min. Keeping the column at temperature 30°C and using the sample amount 10 µL at 5°C and detected all the impurities at 230 nm by the UV detector. In the developed method, elution of Dabigatran was at 26.9 min and all the eluted Impurities were well separated and met the system suitability criteria. The precision is exemplified by relative standard deviation of 0.40%. Method percentage mean recoveries of all the impurities are within the range (90.0% to 115.0%) as per the protocol. Resolution results show that the method is robust. Linearity coefficient for all Impurities is more than 0.999. The LOD obtained is 0.01% and LOQ is 0.03%. A new highly accurate, precise and stability indicating method was developed for related substance of Dabigatran with all the impurities well separate from the degradation product of Dabigatran. It is ready to use for the routine analysis of related substance of Dabigatran in Pharmaceutical industry.