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Combinative application of pH-zone-refining and conventional high-speed counter-current chromatography for preparative separation of alkaloids from Stephania kwangsiensis

Author:
Dong, Hongjing, Zhang, Yongqing, Fang, Lei, Duan, Wenjuan, Wang, Xiao, Huang, Luqi
Source:
Journal of chromatography 2011 v.879 no.13-14 pp. 945-949
ISSN:
1570-0232
Subject:
Stephania, alkaloids, chemical structure, countercurrent chromatography, electrospray ionization mass spectrometry, high performance liquid chromatography, hydrochloric acid, nuclear magnetic resonance spectroscopy, solvents, triethylamine
Abstract:
A method which involves the combination of pH-zone-refining counter-current chromatography (pH-zone-refining CCC) and conventional high-speed counter-current chromatography (HSCCC) was established for the preparative separation of alkaloids from the crude extracts of Stephania kwangsiensis. pH-zone-refining CCC was first performed with the solvent system composed of n-hexane–ethyl acetate–methanol–water (3:7:1:9, v/v), where triethylamine (10mM) was added to the upper organic stationary phase as a retainer and hydrochloric acid (5mM) to the aqueous mobile phase as an eluter. From 2.0g of crude extract, 370mg of sinoacutine and 600mg of a mixture of three other alkaloids were obtained. Then, the mixture was further separated by conventional HSCCC with the solvent system composed of n-hexane–ethyl acetate–methanol–water (7:3:6:4, v/v), yielding 42mg of (−)-crebanine, 50mg of (−)-stephanine and 30mg of l-romerine from 150mg mixture of three other alkaloids, respectively. The purities of the four compounds were all over 98% as determined by HPLC, and the chemical structures of the four compounds were confirmed by positive ESI–MS and ¹H NMR data. Results of the present study successfully indicated that this method was efficient for the preparative separation of alkaloids from natural plants.
Agid:
1003475