Solvent gradient elution for comprehensive separation of constituents with wide range of polarity in Apocynum venetum leaves by high-speed counter-current chromatography.
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.
Zhang
Yuchi
Y
Central Laboratory, Changchun Normal University, Changchun, P. R. China.
Liu
Chunming
C
Zhang
Zhengkun
Z
Qi
Yanjuan
Y
Wu
Guimei
G
Li
Sainan
S
D030010:Apocynum / Q000033:anatomy & histology / Q000737:chemistry; D003377:Countercurrent Distribution / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D015394:Molecular Structure; D018515:Plant Leaves / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry*
D012997:Solvents