title: Combinative application of pH-zone-refining and conventional high-speed counter-current chromatography for preparative separation of alkaloids from Stephania kwangsiensis.
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 (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 eluter. From 2.0 g of crude extract, 370 mg of sinoacutine and 600 mg 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 42 mg of (-)-crebanine, 50 mg of (-)-stephanine and 30 mg of l-romerine from 150 mg 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 (1)H NMR data. Results of the present study successfully indicated that this method was efficient for the preparative separation of alkaloids from natural plants.
authors: Shandong Analysis and Test Center, Shandong Academy of Sciences, Jinan, Shandong, China.
authors: Hongjing
authors: H
authors: Dong
authors: Yongqing
authors: Y
authors: Zhang
authors: Lei
authors: L
authors: Fang
authors: Wenjuan
authors: W
authors: Duan
authors: Xiao
authors: X
authors: Wang
authors: Luqi
authors: L
authors: Huang
mesh_terms: D000085:Acetates; D001060:Aporphines / Q000737:chemistry / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution / Q000379:methods*; D006586:Hexanes; D006863:Hydrogen-Ion Concentration; D000432:Methanol; D009019:Morphinans / Q000737:chemistry / Q000302:isolation & purification*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010936:Plant Extracts / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D031607:Stephania / Q000737:chemistry*; D014867:Water
chemical_list: D000085:Acetates; D001060:Aporphines; D006586:Hexanes; D009019:Morphinans; D010936:Plant Extracts; D014867:Water; C061009:crebanine; C026385:n-hexane; C054160:stephanine; C007650:ethyl acetate; C009270:salutaridine; D000432:Methanol