title: Combined application of macroporous resin and high speed counter-current chromatography for preparative separation of three flavonoid triglycosides from the leaves of Actinidia valvata Dunn.
abstract: In this paper, the combined techniques of macroporous resin column chromatography and high speed counter-current chromatography were applied for preparative separation of flavonoid triglycosides from the leaves of Actinidia valvata Dunn, a famous Chinese medicinal herb. Twelve kinds of macroporous resins were investigated by adsorption and desorption tests. HPD-300 resin showed the maximum effectiveness and thus was selected for the first cleaning-up, in which 20% ethanol was used to remove the undesired constituents and 60% ethanol to elute the targets. The crude extract was then purified by high speed counter-current chromatography with the solvent system composed of ethyl acetate-n-butanol-water (2:1:3 and 4:1:5, v/v). Three flavonoid triglycosides, namely, kaempferol 3-O-α-L-rhamnopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→6)-β-D-galactopyranoside, kaempferol 3-O-α-L-rhamnopyranosyl-(1→3)-(4-O-acetyl-α-L-rhamnopyranosyl)-(1→6)-β-D-galactopyranoside and kaempferol 3-O-α-L-rhamnopyranosyl-(1→3)-(2,4-di-O-acetyl-α-L-rhamnopyranosyl)-(1→6)-β-D-galactopyranoside, were obtained. The purities of the separated compounds were all over 95% as determined by HPLC area normalization method. Their chemical structures were confirmed by UV, MS, NMR, and the standards.
authors.affiliation: Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University, Shanghai, P.R. China.
authors.forename: Liping
authors.initials: L
authors.lastname: Qu
authors.forename: Hailiang
authors.initials: H
authors.lastname: Xin
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Su
authors.forename: Guoyin
authors.initials: G
authors.lastname: Zheng
authors.forename: Changquan
authors.initials: C
authors.lastname: Ling
mesh_terms: D029042:Actinidia / Q000737:chemistry*; D000327:Adsorption; D003377:Countercurrent Distribution / Q000295:instrumentation / Q000379:methods*; D005419:Flavonoids / Q000737:chemistry / Q000302:isolation & purification*; D006027:Glycosides / Q000737:chemistry / Q000302:isolation & purification*; D018515:Plant Leaves / Q000737:chemistry*; D016062:Porosity; D012117:Resins, Synthetic / Q000737:chemistry*
chemical_list: D005419:Flavonoids; D006027:Glycosides; D012117:Resins, Synthetic