To develop and validate a simple, accurate & precise Spectrophotometry methods for simultaneous estimation of Gatifloxacin (Gati) and Loteprednol (Lote) in their combined pharmaceutical dosage form. Two simple, accurate, precise U.V Spectroscopy methods have been developed. First method was based on Simultaneous equation method. Here 286 nm was selected for the estimation of Gatifloxacin and 220 nm was selected for the estimation of Loteprednol. Second method was developed was second order derivative method, here Gatifloxacin was measured at 265.60nm ZCP of Loteprednol and 228.84 was selected for Loteprednol at ZCP of Gatifloxacin. Gatifloxacin and Loteprednol showed linearity in the range of 3-15μg/ml and 5- 25μg/ml in Simultaneous Equation method as well for second order derivative method. Correlation coefficient for Simultaneous estimation method is 0.9936 & 0.9968 whereas for Second order derivative it is 0.9986 & 0.9977 Both methods were validated by validation parameters and it show result where lie within its acceptance criteria as per ICH Q2 (R1) guideline. Hence, it can be successfully used for the routine analysis of Gatifloxacin and Loteprednol in their combined pharmaceutical dosage forms.