A Stability Indicating Reverse Phase High Performance Liquid Chromatography Method for Simultaneous Estimation of Hydroquinone, Hydrocortisone and Tretenoin in Cream Formulation

Abstract

Objective: A simple, sensitive, rapid, precise and accurate stability indicating RP-HPLC method has been developed for simultaneous estimation of Hydroquinone, Hydrocortisone and Tretenoin from their Cream Formulation. Method: The Chromatographic separation was achieved on a reversed-phase Inertsil C18 (4.6 mm I.D. × 250 mm, 5 µm) column using a mobile phase consisting of Buffer (pH 4.0) 0.05M potassium dihydrogen ortho phosphate-Methanol in the ratio of 80:20% V/V at a flow rate of 1ml/min and UV detection at λmax 265 nm. The method showed linearity with correlation coefficient of Hydroquinone, Hydrocortisone and Tretenoin was 0.998, 0.998 and 0.996 over the range of 40-120 µg/ml, 20-60 µg/ml and 0.25-0.75 µg/ml respectively. Result: The retention time was 3 min, 5 min and 6 min for Hydroquinone, Hydrocortisone and Tretenoin respectively. The mean recoveries were found to be in the range of 97.00 –101.00 % for all the components. The method was validated as per the ICH guidelines. The method was validated as per the ICH guidelines. Conclusion: 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.