Simultaneous estimation of biomarkers in hydroalcoholic tuber extract of Amorphophallus paeoniifolius by a validated instrumental thin-layer chromatography method

A plant with potential, Amorphophallus paeoniifolius, which has many medicinal values, belongs to the Araceae family. An accurate and precise method for the quantification of two biomarkers in the tuber extract of A. paeoniifolius has been developed and validated using instrumental thin-layer chromatography (TLC). The planar chromatographic method was developed by utilizing a pre-coated silica gel G60 F₂₅₄ TLC plate as the stationary phase and toluene–ethyl acetate (7:3, V/V) as the mobile phase. For densitometric scanning, 305 nm was chosen as the analytical wavelength for resveratrol. After derivatizing with anisaldehyde‒sulphuric acid, a wavelength of 662 nm was selected to determine β-sitosterol. The optimized conditions showed good separation for both biomarkers, resveratrol and β-sitosterol, with a retention factor (R_F) value of 0.24 and 0.60, respectively. Linear response was obtained in a concentration range of 100–1000 ng/band for both biomarkers. The limits of detection were found to be 10.6 ng/band and 11.8 ng/band for resveratrol and β-sitosterol, respectively, whereas, the limits of quantification were found to be 32.2 ng/band and 35.9 ng/band for resveratrol and β-sitosterol, respectively. The developed method was found to be reproducible, robust, precise, accurate and specific for the evaluation of resveratrol and β-sitosterol in bulk, as well as for the hydroalcoholic tuber extract of A. paeoniifolius.