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Abstract

METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATIONOFHYDROQUINONE, OXYBENZONE AND OCTINOXATE IN SUNSCREEN CREAM BY REVERSED PHASE ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY

Ashok Kumar*, Umme Rab, Vijay Kumar Munipalli, Sujata S. Kaisare, Sayali Warde and Raman Mohan Singh

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Abstract

A simple, precise, accurate, selective, and economic reversed-phase RP-HPLC method has been developed for simultaneous estimation of Hydroquinone, Oxybenzone & Octinoxate in cream formulation. The separation of the drugs was achieved on ACQUITY UPLC@BEH-C18 Column (100mm x 2.1 mm x 1.7µm) using 0.1% Trifluoro acetic acid: Acetonitrile in the ratio of 60:40% v/v as mobile phase. The flow rate was 0.4ml/min and effluents were monitored at 279nm.The method was successfully applied on formulation and there is no interference of matrix components. The linearity of method was obtained in the range of 10µg/ml to 60µg/ml, 5µg/ml to 60µg/ml and 10µg/ml to 75µg/ml of Hydroquinone, Oxybenzone and Octinoxate respectively. Mean percentage recoveries were 100.00% for Hydroquinone and 100.05% for Oxybenzone and 100.04% for Octinoxate. The LOD of Hydroquinone, Oxybenzone and Octinoxate was found to be 3μg/mL, 2μg/mL and 5μg/mL where as the LOQ was 10μg/mL, 6μg/mL and 15μg/mL respectively. The assay values of all true analytes were found to be 99.50%, 99.33% and 99.40% for Hydroquinone, Oxybenzone and Octinoxate respectively. Percentage relative standard deviation of percent assay values for replicate sample preparation was 0.44% for Hydroquinone, 0.44% for Oxybenzone and 0.34% for Octinoxate. The method was robust with respect to the change in flow rate, and composition of the mobile phase. The validation parameters were done in accordance with International Conference on Harmonization (ICH) guidelines. The validation results from statistical analysis of the data, demonstrated the method is reliable.

Keywords: RP-UPLC, Hydroquinone, Oxybenzone, Octinoxate, Brite Cream.


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