HPTLC Method Development and Validation of Genistein and Daidzein Estimation in Mucuna cochinchinensis Seed Extracts

Abstract

Aim: The current study was designed with the objective to estimate the concentration of Genistein and Daidzein in ethyl acetate extract of Mucuna cochinchinensis by high-performance thin-layer chromatography (HPTLC).Methods: The separation of bioactive compounds was performed using mobile phase, Toluene: n-Butanol: Glacial Acetic Acid: Water (1:4:2:2) and detected at wavelength 254 nm. The method was validated for linearity, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ) by the International Conference on Harmonization guidelines (ICH). Results: The calibration range was found to be 10–50 and 20–100 ng/band for Genistein and Daidzein, respectively. Daidzein was separated with a R_f value of 0.34 and Genistein with a R_f value of 0.47. Average recovery was 99.96% and 99.90% for Genistein and Daidzein, respectively. The LOD and LOQ were 0.89 ng and 2.98 ng for Genistein, and 5.1 ng and 17.29 ng for Daidzein, respectively. Conclusion: 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.

Keywords: Daidzein, Genistein, high-performance thin-layer chromatography (HPTLC), Isoflavones, M. cochinchinensis, method validation

Cite this Article

K. Natarajan, S. Sangeetha, Vineeth Chandy. HPTLC Method Development and Validation of Genistein and Daidzein Estimation in Mucuna cochinchinensis Seed Extracts. Research & Reviews: A Journal of Drug Formulation, Development and Production. 2020; 7(1): 16–25p.