The present study was aimed to develop a simple and validated chromatographic method for the estimation of Trimyristin in the API and in poly herbal Ayurvedic formulations by UFLC method, along with stress induced degradation studies of the drug and validate the method as per the ICH guidelines. The analysis was carried out under isocratic conditions using dichloromethane and acetonitrile in the ratio of 95:5 as mobile phase with a flow rate of 1.0 ml/min at 210nm. The method was simple, specific, precise, and accurate with retention times 0.904 and 0.906 minutes for the API and isolated product. The system obeys the Beer's law in the concentration range of 10-90μg/ml. The percentage recovery studies performed showed a percentage recovery of 98.3 -101.6%w/v and found to be linear with a correlation coefficient (r2) of 0.9993. The method was found to be precise with relative standard deviation of less than 2%, detection limit and quantitation limit was estimated to be 4.58μg/ml, 15.27μg/ml respectively. The method was found to be robust even by change in mobile phase ratio of ±5%, change in wavelength and change in flow rate of ±0.1 ml/min. This validated method was sensitive and reproducible enough to be used for routine analysis of Poweromin tablet and Makaradhwaj vatti in time and cost effective manner.