A simple, economic, selective, precise, and accurate High Performance liquid Chromatographic method for the analysis of Ametoctradin in bulk and commercial formulations was developed and validated in the present study. The mobile phase consists of a mixture of Methanol and water in the proportion 70:30 and adjust the pH to 6.0 ± 0.05 with sodium hydroxide solution. This was found to give a sharp peak of Ametoctradin at a retention time of 7.865 min. HPLC analysis of Ametoctradin was carried out at a wavelength of 294 nm with a flow rate of 1.0 ml/min. The linear regression analysis data for the calibration curve showed a good linear relationship with a regression coefficient of 0.999 in the concentration range of 50 µg ml-1 to 150 µg ml-1. The linear regression equation was y =2160x-495.9. The developed method was employed with a high degree of precision and degradation for the analysis of Ametoctradin. The developed method was validated for precision, robustness, detection and quantification limits as per the ICH guidelines. The wide linearity range, sensitivity, short retention time and composition of the mobile phase indicated that this method is better for the quantification of Ametoctradin.