Design of Experiment Based Validated HPTLC Method for Analysis of Deflazacort in Tablet Formulation

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. The samples were spotted in the form of bands of width 5 mm with a Camag microlitre syringe on pre-coated silica gel aluminum plate 60 F254, mobile phase was composed of toluene: methanol: glacial acetic acid (8.3: 1.2: 0.5 v/v/v). For the same CMPs selected were organic phase (toluene: methanol), mobile phase volume, saturation time, band width, solvent front, wavelength, type of chamber and CAA was identified as retention factor21 Factor screening study: A seven factor eight -run Fractional factorial design was used for factor screening study to identify the CMPs affecting CAA's (i.e. Rf ). A standard concentration 480ng/band was used for all experimental runs, which were analyzed for CAAs namely peak area and retention factor (Table II) Optimization data analysis and model validation: