ABSTRACT: In the current study, a reverse phase high performance liquid chromatography (RP-HPLC) approach that was developed and validated for the simultaneous quantification of hydroquinone, allantoin, and tretenoin in a cream formulation is discussed. On a reversedphase InertsilC18 column (4.6 mm I.D. 250 mm, 5 m), the chromatographic separation was completed using a mobile phase made up of buffer (pH 3.5), 0.05 M potassium dihydrogen ortho phosphate-methanol (70:30% V/V), and UV detection at λmax 223nm. The technique demonstrated linearity, with Hydroquinone, Allantoin, and Tretenoin correlation coefficients of 0.999, 0.999, and 0.999 spanning ranges of 100-300 g/mL, 50 to 150 g/mL, and 0.625 to 1.875 g/mL, respectively. All of the components' mean recoveries ranged from 99.00 to 101.00%. The technique was approved in accordance with ICH standards. With no unwelcome interference, the established approach was extremely precise, repeatable, and detectable for hydroquinone, allantoin, and tretenoin together. The approach is effective in isolating the API from its degradants when judged on many criteria, including system suitability, precision, accuracy, linearity, robustness, and stability studies. It can be used to analyse samples of hydroquinone, allantoin, and tretenoin.