Abstract

DEVELOPMENT OF ANALYTICAL METHOD FOR SIMULTANEOUS ESTIMATION OF HYDROQUINONE AND MONOBENZONE IN TOPICAL FORMULATION BY RP-HPLC

Harikishor Barange* and Suhail Asghar

Abstract

A new specific, precise, accurate and robust RP-HPLC method has been developed for the simultaneous determination of Hydroquinone and Monobenzone in a pharmaceutical cream formulation. The chromatographic separation was carried out at Analytical technology HPLC instrument (Software: HPLC Work Station) equipped with Deuterium lamp as detector, HPLC pump and manual injecting facility programmed at 20 μL capacity per injection was used. The stationary phase was Zodiac Column C18 (250 mm x 4.6 mm, 5μm) at ambient temperature. The mobile phase was Acetonitrile: Triethylamaine Buffer (60:40) adjust pH 3.5 by 10% orthophosphoric acid. Detection was carried out at 230nm using UV Detector. The flow rate was 1.0mL/min and retention time was about 3.3mins and 5.1mins for Hydroquinone and Monobenzone. The linearity was obtained in the concentration range of 24-64μg/mL and 30-70μg/mL for Hydroquinone and Monobenzone respectively. Mean percentage recoveries were 99.89% for Hydroquinone and 99.57% for Monobenzone. The LOD of Hydroquinone and Monobenzone was found to be 1 μg/mL and 2.0μg/mL whereas the LOQ was 5μg/mL and 10 μg/mL respectively. The assay values of both the analytes was found to be well within the limits that is 100.05% and 99.55% for Hydroquinone and Monobenzone respectively. Percentage relative standard deviation of percent assay values for replicate sample preparation was 1.09% for Hydroquinone and 0.96% for Monobenzone. The method was robust with respect to change in flow rate, and composition of mobile phase.

Keywords: Cream formulation, Hydroquinone, Monobenzone, Simultaneous Estimation, RP-HPLC analytical method development, Validation.