A simple, rapid, precise, sensitive and reproducible reverse phase high performance liquid chromatography (RP-HPLC) method has been developed for the quantitative analysis of Mitapivat in pharmaceutical dosage form. Chromatographic separation of Mitapivat was achieved on Waters Alliance-e2695, by using Zorbax SB C18 (250x4.6mm, 5µ) column and the mobile phase containing ACN and Water in the ratio of 80:20% v/v. The flow rate was 1.0 ml/min; detection was carried out by absorption at 278nm using a photodiode array detector at ambient temperature. The number of theoretical plates and tailing factor for Mitapivat were NLT 2000 and should not more than 2 respectively. %Relative standard deviation of peak areas of all measurements always less than 2.0. The proposed method was validated according to ICH guidelines. The method was found to be simple, economical, suitable, precise, accurate & robust method for quantitative analysis of Mitapivat. Key words: HPLC Method, Mitapivat, ICH guidelines, Validation, Degradation studies.