A new, selective and accurate spectrofluorimetric assay has been described for detection of Amisulpride and Memantine hydrochloride in pharmaceutical formulations and real plasma samples. The described assay depends on the reaction between the primary amino group of the selected drugs with acetyl acetone & formaldehyde in an acetate buffer of pH 4.8. The derivatized product showed yellow fluorescence at ex= 418 nm and em= 484.5 nm. The calibration graph was linear in the range of 0.05-0.5 and 0.2-1 µg mL1 for AMS and ME, orderly. The limits of detection were 0.0085 and 0.0153 µg mL1, and the limits of quantitation were 0.026 and 0.0464 µg mL1 for AMS and ME respectively. Validation of the described assay was in consonance with ICH guideline. Due to the sensitivity of the prescribed assay, it permits the determination of selected medications in biological sample quantitatively.