A Stability indicating Reverse Phase High Performance Liquid Chromatography Method for simultaneous estimation of Allantoin, Hydroquinone and Tretenoin in Cream formulation

Abstract

A simple, sensitive, rapid, precise and accurate RP-HPLC method has been developed for simultaneous estimation of Hydroquinone, Allantoin and Tretenoin from Cream Formulation. The Chromatographic separation was achieved on a reversed-phase InertsilC18 (4.6mm I.D. × 250mm, 5µm) column using a mobile phase consisting of Buffer (pH 3.5) 0.05M Potassium dihydrogen ortho phosphate-Methanol (70:30% V/V) at a flow rate of 1mL/min and UV detection at λmax 223 nm. The method showed linearity with correlation coefficient of Hydroquinone, Allantoin and Tretenoin was 0.999, 0.999 and 0.999 over the range of 100-300 µg/mL, 50-150 µg/mL and 0.625-1.875 µg/mL respectively. The mean recoveries were found to be 99.00 to 101.00 % for all the components. The method was validated as per the ICH guidelines. The method was stable and specific and when sample was stressed under different conditions like Acid, Base, Oxidative, Thermal and Photolytic, no interference of degradants were observed.