The current work studied the stability of neomycin sulfate (NS) in its pharmaceutical dosage form. The ability of NS to withstand alkaline, acidic, and oxidative forced degradation conditions was evaluated. We developed a RP-HPLC method over a Kromasil C18(w) using a mobile phase of 0.1% aqueous phosphoric acid maintained at a flow rate of 1mL/min within a relatively short run time of 3.0 min. The UV-detector monitored the effluent at 210.0 nm, NS expressed a correlation coefficient approaching unity within the concentration range 40-200 µg/mL. The method achieved adequate specificity for NS in presence of the products of degradation and excipients of the dosage form. Validation parameter were calculated according to the USP guidelines. The stability study expressed poor NS stability in either of the three degradation conditions. The procedure fulfilled the greenness criteria of the four quadrants of the National Environmental Methods Index metric. The method represents a reliable green analytical solution for NS stability assessment in quality control laboratories.