Abstract

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Two sensitive, selective and precise chromatographic methods have been developed, optimized and validated for Allopurinol (ALP) and Benzbromarone (BENZ) determination in their pure form, laboratory prepared mixtures and pharmaceutical dosage form. The first developed method was based on thin layer chromatographic (TLC) combined with densitometric determination of the separated spots. The separation was achieved using silica gel 60F254 TLC plates. The mobile phase used was chloroform: methanol (9.2:0.8, v/v) and UV detection at 240nm.Good correlations were obtained between the integrated peak area of the studied drugs and their corresponding concentrations in different ranges. The second developed method was based on the high-performance liquid chromatography with ultraviolet detection, by which the proposed components were separated using Zorbax C18 column using a mobile phase consisting of sodium acetate buffer (pH=4.5, adjusted with acetic acid): acetonitrile: triethylamine (50:50:0.5, by volume) maintaining the mobile phase flow rate at 1mLmin-1with UV detection at 260nm. Different parameters affecting the suggested methods were optimized for maximum separation of the cited components. System suitability parameters of the two developed methods were also tested. Validation of the methods has been carried out according to USP requirements and ICH guidelines, accuracy, precision and repeatability were found to be within the acceptable limits. The results obtained by TLC-Densitometric and RP-HPLC methods were statistically compared with those obtained by the reported RP-HPLC method and no significant difference was found regarding both accuracy and precision.

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