In this study, a simple and sensitive fluorimetric method was described for the determination of Ascorbic Acid (AA). The procedure is based on the reaction between AA and Methylene Blue (MB). The fluorescence intensity of NIB was measured at excitation and emission of 664 and 682 nm, respectively. MB concentration was decreased as a function of decreasing fluorescence intensity due to forming colorless form of MB (Leuco-MB) in the reaction between AA and NIB. A linear relationship was obtained between the decreasing fluorescence intensity and the concentration of AA in the range of 3.0x10(-7)-6.0x10(-6) mol(.)l(-1). The detection limit was 2.5x10(-7) mol(.)l(-1). The proposed method was applied successfully for the determination of AA in Vitamin C tablets.