A novel reverse phase Ultra performance liquid chromatographic technique was developed for the determination of balofloxacin in bulk and pharmaceutical dosage forms. The method was developed using waters Acquity BEH 50mm, 2.1mm, 2μm, C 18 column with mobile phase containing a gradient mixture of 0.1% phosphoric acid and acetonitrile. Detection was carried out at wavelength 295 nm. The retention time of balofloxacin was 0.89 min. The method showed good linearity in the range 0.5, 1, 1.5,2,3 µg/ml with correlation coefficient for balofloxacin. The proposed method has been validated as per ICH guidelines and successfully applied to the estimation of balofloxacin in their tablet dosage form.