A simple, accurate and precise High Performance Liquid Chromatographic (HPLC) method has been developed for simultaneous determination of Gatifloxacin and Loteprednol in bulk and dosage form. The method has been validated as per the guidelines of ICH. The separation is achieved on 250*4.6mm C18, 5micron (Hypersil BDS) column with flow rate 1.0 mL per minute in isocratic mode using Buffer pH3.5: Acetonitrile (45:55) as mobile phase. Column oven temperature is maintained at 25°C and observations are recorded at 275 nm. The linearity range was found to be in the range of 15-45 μg/ml for Gatifloxacin and 25-75 μg/ml for Loteprednol. Correlation coefficient for calibration curve of Gatifloxacin and Loteprednol was found to be 0.9977 and 0.9983 respectively. The method is simple, accurate, reproducible and short and can be used for simultaneous analysis of Loteprednol and Gatifloxacin.