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Solvent gradient elution for comprehensive separation of constituents with wide range of polarity in Apocynum venetum leaves by high‐speed counter‐current chromatography
Author(s) -
Zhang Yuchi,
Liu Chunming,
Zhang Zhengkun,
Qi Yanjuan,
Wu Guimei,
Li Sainan
Publication year - 2010
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201000308
Subject(s) - chemistry , chromatography , ethyl acetate , countercurrent chromatography , elution , methanol , column chromatography , organic chemistry
A novel gradient elution was efficiently utilized for the separation of the chemical components with a wide range of polarity from the mixed extract of the Chinese medicinal herb Apocynum venetum or mixed standards by high‐speed counter‐current chromatography. Three sets of solvent systems, n ‐hexane–ethyl acetate–methanol–water (1.5:3.5:2:4.5 v/v/v/v), ethyl acetate–methanol–water (5:2:5 v/v/v) and n ‐butanol–methanol–water (5:1:5 v/v/v) were used for the one‐step elution. The separation was initiated by filling the column with the lower phase of n ‐hexane–ethyl acetate–methanol–water (1.5:3.5:2:4.5 v/v/v/v) as a stationary phase followed by elution with the upper phase of n ‐hexane–ethyl acetate–methanol–water (1.5:3.5:2:4.5 v/v/v/v) to separate the hydrophobic compounds (tail to head). Then the mobile phase was switched to the upper phase of ethyl acetate–methanol–water (5:2:5 v/v/v) to elute the moderate hydrophobic compounds, and finally the hydrophilic compounds still retained in the column were fractionated by eluting the column with the upper phase of n ‐butanol–methanol–water (5:1:5 v/v/v). A total of 13 compounds including adhyperforin, hyperforin, amentoflavone, biapigenin, quercetin, astragalin, trifolin, isoquercetin, hyperside, acetyled hyperside, rutin, chlorogenic acid and quercetin‐3‐ O ‐β‐ D ‐glucosyl‐β‐ D ‐glucopyranoside were successfully separated via the three sets of solvent systems in one‐step operation for 90 min.