This research included the study of different factors that may effect on gatifloxacin stability (anew quinolone synthetic antibacterial agent) in its aqueous solution in order to develop and optimize the best delivary of the drug to the eye (as eye drop) with maximum local concentration and minimum systemic absorption and toxicity.Different formulas of gatifloxacin solution for ophthalmic use (0.3%)w/v were prepared in citrate, acetate,citrate/phosphate and phosphate buffers,their tonicity adjusted with suitable quantity of sodium chloride.The effect of different factors that might affectthe stability of gatifloxacin in its prepared ophthalmic solution was studied and determined spectrophotometrically at 287 nm. The results showed that The use of disodium edetate as asequestering agent gave more stable formula and gatifloxacin undergoes hydrolysis at low pH with optimum stability at pH 6.0, which is the most suitable pH for this ophthalmic solution. The type of buffer significantlyaffects on the rate of hydrolysis of gatifloxacinspecially at low pH and optimum stability was obtained by using phosphate buffer. The concentration of phosphate buffer had a significant effect on the hydrolysis of gatifloxacin and the rate of hydrolysis increased as the concentration buffer increased. Ionic strength affects the hydrolysis rate of gatifloxacin and the hydrolysis increased as the ionic strength increased. Light had a significant effect on the rate of hydrolysis of the drug and the drug losses 10% of its potency after 10 monthes of light exposure at room temperature. The prepared formula J ( gatifloxacin 0.3% in 0.1M phosphate buffer with sodium chloride 0.26% , xanthan gum 0.2% and disodium edetate 0.01%) is thebest stable one and had no irritation on the eye of experimental animals, and it passes successfully quality control tests including: drug content, pH, clarity and sterility test and comply with united state pharmacopoeia for ophthalmic solutions.