Application of HPTLC, spectrofluorimetry and differential pulse voltammetry for determination of the antifungal drug posaconazole in suspension dosage form

Abstract

This work presents the development, validation and application of three simple and direct analytical methods for determination of posaconazole (PSZ) in its pure form and in suspension dosage form. Method I is based on high performance thin layer chromatography (HPTLC) where effective separation of PSZ and the internal standard (itraconazole) was achieved using Merck HPTLC plates (20×10cm aluminium plates with 250μm layer thickness precoated with silicagel 60 F254) and a mobile phase composed of acetone and chloroform (1:2, by volume), followed by densitometric measurement of the drugs' spots at 262nm. Method II involves measurement of the native fluorescence of PSZ in 0.1M H2SO4 at excitation and emission wavelengths of 260 and 365nm, respectively. Method III depends on the voltammetric analysis of PSZ. A well-defined cathodic wave was obtained for PSZ in Britton-Robinson buffer pH 6.5 using the differential-pulse mode at the hanging mercury drop electrode (HMDE). The developed methods were validated according to the International Conference on Harmonization (ICH) guidelines regarding linearity, ranges, accuracy, precision, robustness and limits of detection and quantification. The proposed methods showed good linearity over the concentration ranges 5-50, 0.05-0.3, 0.005-0.05μg/mL PSZ for methods I, II, and III respectively. Intra and inter-day precision were verified by the RSD% values which were less than 2%. The proposed methods were successfully applied for the quantification of PSZ in suspension dosage form with no observable interferences. Assay methods were favorably compared with those obtained by previously reported HPLC method.