Development and Validation of Stability Indicating RP-HPLC Method for Simultaneous Determination of Anti-Haemorrhagic Ethamsylate and Anti-Fibrinolytic Tranexamic Acid in Combined Formulation using Green Assessment

Abstract

The present study demonstrates the method development and validation for simultaneous estimation of Ethamsylate & Tranexamic acid in combined pharmaceutical dosage form by using RP-HPLC along with the stability studies. According to ICH guidelines for stability testing of new drug substances and products, both the drugs were subjected to various stress conditions. The chromatographic separation was carried out on ALTIMA C18 column with the dimensions of 150 mm x 4.6 mm x 5µm using Waters 2695 HPLC instrument equipped with 2998 series of PDA detector. The isocratic mobile phase used was made up of Phosphate Buffer with pH adjusted at 3.1 & Acetonitrile (ACN) in the ratio 80:20 v/v with the flow rate of 1.0 ml /min and both the drugs have been detected using UV detector at 249 nm. At room temperature Ethamsylate and Tranexamic acid were found to have retention times of 2.353 & 3.033 minutes respectively with a total run time of 6 minutes. The developed method was found to be linear at the concentration range of 25 µg/ml – 150 µg/ml with coefficient of determination observed at 0.09993 for Ethamsylate & 0.9998 for Tranexamic acid respectively. The developed method was validated by utilizing various validation parameters and the force degradation & stability studies were applied to analyse the stability and to identify settlement of the degradation products. The developed method was also analysed using two tools like AGREE and GAPI for assessing the green profile of the developed method. The established method can be applied for the simultaneous determination of ETS & TXA in combined dosage form.