A new, simple, sensitive, precise and robust high-performance thin layer chromatographic (HPTLC) method was developed for the estimation of Tribulosin and (Mono ammonium glycyrrhizinate) MAG in herbal extracts and polyherbal formulation. Analysis of Tribulosin and MAG were performed on TLC aluminium pre-coated plates with silica gel 60F-254 as stationary phase. Linear ascending development was carried out in twin trough glass chamber saturated with mobile phase consisting of butanol: GAA: water (6: 1: 3, v/v/v) for Tribulosin and chloroform: methanol: GAA: water: formic acid (1.8: 0.4: 0.8: 0.2: 0.1, v/v/v/v/v) for MAG at room temperature (25±2°C). After development the plate was derivatized with vanillin-sulphuric acid (H2SO4) for both Tribulosin and MAG. Camag TLC scanner III was used for spectrodensitometric scanning and analysis of plate in absorbance mode at 454 and 254 nm for Tribulosin and MAG respectively. The system was found to give compact spots for Tribulosin (Rf 0.41) and MAG (Rf 0.56). The data for calibration plots showed good linear relationship with r2=0.9994 and r2=0.9992 in the concentration range of 75–450 ng and 40-240 ng for Tribulosin and MAG respectively. According to international conference on harmonization (ICH) guidelines the present method was validated for precision, repeatability and recovery. The limits of detection and quantification were determined. Statistical analysis of the data showed that the method is reproducible and selective for estimation of Tribulosin and MAG.
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