Development and Validation of Stability-Indicating RP-HPLC and UV Spectrophotometric Methods for Quantitative Determination of Hydroquinone in Pharmaceutical Formulations

Research Article

Abstract

This research work focusses on two validated analytical methods for the quantification of hydroquinone in pharmaceutical formulations using reverse-phase high-performance liquid chromatography (RP-HPLC) and UV spectrophotometry. The RP-HPLC method utilized an Agilent C18 column (250 mm × 4.6 mm, 5 μm) with a mobile phase consisting of methanol and 0.05% orthophosphoric acid (50:50 v/v, pH 3.7) at a flow rate of 0.7 mL/min. Detection was performed at 290 nm with a retention time of 3.859 minutes. The UV spectrophotometric method was developed using methanol as the solvent, with measurements taken at 290 nm. Both methods demonstrated linearity in their respective ranges: 10-50 μg/mL for RP-HPLC and 1-5 μg/mL for UV spectrophotometry, with correlation coefficients exceeding 0.999. Method validation parameters including accuracy, precision, robustness, and system suitability were evaluated according to ICH guidelines. The limits of detection and quantification for the RP-HPLC method were 0.0147 μg/mL and 1.3930 μg/mL, respectively. Recovery studies showed excellent results ranging from 98.56% to 100.24%. Both methods were successfully applied to commercial tablet formulations with high accuracy and precision. The developed methods were found to be simple, rapid, and reliable for routine quality control analysis of hydroquinone in pharmaceutical preparations.