Development and validation of a high-performance thin-layer chromatographic method for the determination of biomarker β-Amyrin in the leaves of different Ficus species

A simple and sensitive high-performance thin-layer chromatographic (HPTLC) method was developed for the evaluation of biomarker β-Amyrin in the leaves of five different species of genus Ficus (Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata, and Ficus vasta) grown in the Kingdom of Saudi Arabia. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with solvents toluene-methanol (9:1, v/v) as the mobile phase. After development, the HPTLC plate was derivatized with p-Anisaldehyde reagent to give well-resolved and compact spot of β-Amyrin. Scanning and quantification were done at 550 nm. The system was found to give compact spot for β-Amyrin at RF = 0.58. The linear regression analysis data for the calibration plots showed good linear relationship with r2 = 0.998 with respect to area in the concentration range of 100-900 ng. The regression equation for β-Amyrin standard was found to be Y = 5.835X + 87. The precisions (n = 6) for β-Amyrin were found to be 1.64-1.77% and 1.68-1.84%, respectively, for intra-day and inter-day batches, and the recovery values were found to be 97.6-98.3%. β-Amyrin was found to be present in three species, i.e., F. carica (0.29%, w/w), F. nitida (0.54% w/w), and F. palmata (0.31%, w/w), while it was absent in F. vasta and F. ingens. The statistical analysis proves that the developed method for the quantification of β-Amyrin is reproducible; hence, it can be employed for the determination of β-Amyrin in plasma and other biological fluids as well as in finished products available in the market.