ABSTRACT:
Deflazacort is an oxazoline derivative of prednisolone with anti-inflammatory and immunosuppressive activity. The present study describes the development of HPTLC method based on systematic design of experiment followed by validation of the method for the analysis of Deflazacort in bulk and tablet dosage form. Systematic optimization was performed employing Box-Behnken design (BBD). Mobile phase ratio, band width, solvent front were selected as the critical method parameters (CMPs) for initial screening studies. Search for optimum chromatographic conditions was carried out with numerical and graphical optimization and the design space was generated with peak area and retention factor as critical analytical attributes (CAAs). Based on the predicted optimized chromatographic condition, aluminium plates pre-coated with silica gel 60 F254 as the stationary phase and toluene: methanol: glacial acetic acid (8.3:1.2:0.5% v/v/v) as the mobile phase. Quantification was achieved based on densitometry analysis of Deflazacort over the concentration range of 120-720 ng/band at 243 nm. The method yielded compact and well-resolved bands at Rf of 0.43 ± 0.03 for Deflazacort. The developed method was validated according to ICH guidelines. Further validated method was used to study degradation of Deflazacort when exposed to stress conditions like acidic and alkaline hydrolysis, oxidative and photolytic degradation. Developed validated method applied for forced degradation studies make the method highly efficient for the routine analysis of marketed tablet formulations.