Two modes of high-speed counter-current chromatography (HSCCC) were successfully applied to the separation of alkaloids from crude extract of Nelumbo nucifera leaves. The conventional HSCCC separations were performed with a two-phase solvent system composed of tetrachloromethane-CHCl3-methanol-0.1 M HCl at a volume ratio of 1:3:3:2 (v/v/v/v), and 120 mg crude extract could be successfully separated. pH-Zone-refining CCC was performed with a two-phase solvent system composed of petroleum ether (60-90 °C)-ethyl acetate-methanol-water (5:5:2:8, v/v/v/v) where triethylamine (10 mM) was added to the upper organic stationary phase as a retainer and hydrochloric acid (5 mM) to the aqueous mobile phase as an eluent. From 4.0 g of the crude extract, 120 mg N-nornuciferine, 1020 mg nuciferine and 96 mg roemerine were obtained in a single run each with a purity of over 98% as determined by HPLC. The structures of the isolated compounds were identified by ESI-MS, 1H NMR and 13C NMR. © 2010 Elsevier B.V. All rights reserved.
Zhenjia, Z., Minglin, W., Daijie, W., Wenjuan, D., Xiao, W., & Chengchao, Z. (2010). Preparative separation of alkaloids from Nelumbo nucifera leaves by conventional and pH-zone-refining counter-current chromatography. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 878(19), 1647–1651. https://doi.org/10.1016/j.jchromb.2010.04.020