A rapid and precise reverse phase high performance liquid chromatographic method has been developed for the validated of Dalfampridine in its pure form as well as in tablet dosage form. Chromatography was carried out on ODS C18 (4.6 x 250mm, 5µm) column using Acetonitrile and water in the ratio of 80:20 v/v,as the mobile phase at a flow rate of 1.0mL/min, The retention time obtained for DFP was 2.98 min. The method produce linear responses in the concentration range of 20 µg/ml of Dalfampridine . The method precision for the determination of assay was below 2.0%RSD. The method is useful in the quality control of bulk and pharmaceutical formulations.