Method Development for the Chromatographic analysis of a Two-Component Tablet Formulation Using Chemometric Optimization Technique

Abstract

A new ultra-performance liquid chromatography (UPLC) method was improved for the determination of sulfamethoxazole and trimethoprim in a two-component tablet formulation. In the development of the UPLC method, the chromatographic conditions were chemometrically optimized by using the experimental design and optimization procedure. The central composite design and fitting model was applied to identify the suitable chromatographic conditions providing a desirable elution of sulfamethoxazole and trimethoprim in a chromatogram. In the central composite design, temperature, flow rate and buffer% were selected as the effective factors on the chromatographic resolution. The buffer system was the mixture of 0.1 M CH3COOH and CH3COONa (pH 4.75) in mobile phase system. In the optimization step, the chromatographic conditions were found to be 0.20 mL/min for flow rate, 38.0 °C for the column temperature and 66% for the acetate buffer system (v/v) in the mobile phase. Separation of the analyzed compounds was performed on a Waters BEH C18 column (100 mm-2.1 mm, 1.7 mm i.d.). In the validation step, recovery study was performed by applying the UPLC method to the synthetic binary mixture of sulfamethoxazole and trimethoprim. Recovery results were found as 99.12% for sulfamethoxazole and 99.44% for trimethoprim. The chemometric optimized UPLC method was applied to the quantitation of sulfamethoxazole and trimethoprim in commercial tablets.