Attempts were made to develop RP-HPLC method for simultaneous estimation of Lefamulin from Tablet. For the RP - Agilent Tech. Gradient System with Auto injector, UV (DAD) & Gradient Detector Reverse Phase (Waters) C18 column (4.6mm x 100mm; 2μm), a 20μl injection loop and UV730D Absorbance detector and running chemstation 10.1 software. Methanol: water (0.05%OPA), (50:50) v/v, pH 3. was used as the mobile phase for the method. The detection wavelength was 278 nm and flow rate was 1 ml/min. In the developed method, the retention time of Lefamulin were found to be be 3.513 min. The developed method was validated according to the ICH guidelines. The linearity, accuracy, precision, range, robustness was within the limits as specified by the ICH guidelines. Hence the method was found to be simple, accurate, precise, economic and reproducible. So the proposed methods can be used for the routine quality control analysis Lefamulin in bulk drug as well as in formulations