Simultaneous estimation of genistein and daidzein in Pueraria tuberosa (Willd.) DC by validated high-performance thin-layer chromatography (HPTLC) densitometry method

The present study was performed to estimate the concentration of genistein and daidzein in ethanol extract of tubers of Pueraria tuberosa (Indian kudzu or Vidarikanda) and its various fractions (n-hexane, ethyl acetate, n-butanol, and aqueous) by high-performance thin-layer chromatography (HPTLC). The separation of bioactive compounds was performed using mobile phase, toluene:ethyl acetate:acetone:formic acid (20.0:4.0:2.0:1.0) and detected at wavelength 269?nm. The method was validated for linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), etc. by International Conference on Harmonization guidelines. The calibration range was found to be 100–600?ng/band for both the bioactive compounds. Daidzein was separated with an R_f value of 0.39?±?0.02 and genistein with an R_f value of 0.54?±?0.02. Average recovery was 99.96 and 99.90% for genistein and daidzein, respectively. The LOD and LOQ were 14.786 and 44.805?ng, respectively, for genistein, and 9.607 and 29.114?ng, respectively, for daidzein. Both the phytoconstituents were found in ethanol extract and its ethyl acetate fraction only. The developed HPTLC method was simple, precise, robust, specific, rapid, and cost effective and could be used for quality control analysis and quantification of genistein and daidzein in different herbal formulations containing the plant species.