Preparative separation of alkaloids from Nelumbo nucifera leaves by conventional and pH-zone-refining counter-current chromatography

College of Food Science and Engineering, Shandong Agricultural University, 61 Daizong Street, Taian, Shandong 271018, China.
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (Impact Factor: 2.73). 06/2010; 878(19):1647-51. DOI: 10.1016/j.jchromb.2010.04.020
Source: PubMed


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 degrees 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.

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