The aim of this research was to develop and validate a spectrophotometric method for determination of tranexamic acid by using derivatization technique. Tranexamic acid is non-chromophoric drug contains no π electrons to act as fluorophore or chromophore so its determination by conventional spectrophotometric method is not possible. So derivatization reaction was performed for determination of tranexamic acid through the reaction with 2, 4- dinitrofluorobenzene (DNFB) as chromogenic derivatizing agent to produce a yellow coloured product which absorbs maximally at wavelength (λmax) 370nm. Beer’s law was obeyed over the concentrations range of 6-16 μg/ml with lower detection limit of 0.3586μg/ml. The proposed method was applied for quantitation of the studied drug in its pure form with mean percentage recoveries of 100.96%, 99.83% and 99.60% for 80, 100 and 120% respectively. Different experimental parameters affecting the development and stability of the coloured were studied and optimized. The suggested method is very sensitive and may have the potential to be used for the analysis of tranexamic acid.