Multiple Marker-Based Evaluation of Kalanchoe pinnata, Bombax ceiba, and Morus alba Leaves: Quantification of alpha-Amyrin, Lupeol, and beta-Sitosterol Using High-Performance Thin-Layer Chromatography

摘要

Kalanchoe pinnata (Crassulaceae), Bombax ceiba (Bombaceae), and Morus alba (Moraceae) are important drugs in Indian and Chinese Systems of Medicine. Leaves of K. pinnata are reported to show anti-inflammatory, antifungal, and antibacterial activity and are reported to contain quercetin, kaempferol, alpha-amyrin, beta-sitosterol, ferulic, and caffeic acid. Leaves of B. ceiba are reported for the treatment of diarrhea, dysentery, gonorrhea, and bladder ulceration and are reported to contain shamimin, lupeol, beta-sitosterol, and alpha-amyrin. Leaves of M. alba are reported to show anti-inflammatory, emollient, hypolipidemic, and diuretic activity and are reported to contain quercetin, rutin, moracetin, and artocarpin. In the present work, a method for simultaneous quantification of three marker compounds, viz., alpha-amyrin, lupeol, and beta-sitosterol from the leaves of K. pinnata, B. ceiba, and M. alba was developed and validated as per International Conference on Harmonization (ICH) guidelines. This is the first report of simultaneous quantification of these three compounds. alpha-Amyrin, lupeol, and beta-sitosterol resolved at R-F 0.71, 0.28, and 0.16, respectively, on thin-layer chromatography (TLC) when developed in the solvent system of toluene-ethyl acetate (9.5:0.5). The linearity range for alpha-amyrin, lupeol, and beta-sitosterol was found to be 160-560, 150-900, and 80-480 ng spot(-1), respectively, with correlation coefficients (r values) of 0.999, 0.997, and 0.995, respectively. The amount of alpha-amyrin, lupeol, and beta-sitosterol were found to be in range of 0.013-0.047, 0.046-0.318, and 0.028-0.123 % w/w, respectively. The developed methods were found to be accurate, precise, and reproducible and can be used for routine quality control of herbal material and formulations containing K. pinnata, B. ceiba, and M. alba.