Solvent gradient elution for comprehensive separation of constituents with wide range of polarity in Apocynum venetum leaves by high-speed counter-current chromatography
✍ Scribed by Yuchi Zhang; Chunming Liu; Zhengkun Zhang; Yanjuan Qi; Guimei Wu; Sainan Li
- Publisher
- John Wiley and Sons
- Year
- 2010
- Tongue
- English
- Weight
- 194 KB
- Volume
- 33
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
Abstract
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.