A high performance liquid chromatography method has been developed for determination of letrozole in tablets. The method has developed in reverse phase column with acetonitrile-50.0 mM phosphoric acid solution, pH:7 (50:50, v/v) and 0.7 mL/min flow rate by using a fluorimetric detector. Letrozole has natural fluorescence properties, and there is no need for derivatization for this molecule. The detector was set at 256, 585 nm for excitation and emission wavelength respectively. The method was validated in accuracy, precision, specificity, linearity and system suitability parameters are studied. The linear range was found 50-700 ng/mL. The limit of detection and quantification for letrozole was found to be 14-50 ng/mL respectively. The recovery of drug is found to be 98.8%. The developed method has been validated and successfully applied to tablet analysis of drug substance. This method is simple, reproducible and can be used safety for routine analysis of letrozole in tablets.