Comprehensive separation of major compositions from Sophora japonica var. violacea by counter-current chromatography using a liquid-liquid extraction strategy

Bioassay-guided fractionation of the ethanol extract of whole herbs of Achillea alpina led to the isolation of isochlorogenic acids A and B as transient receptor potential vanilloid 3 (TRPV3) channel antagonists by using a calcium fluorescent assay. The structures were identified by spectroscopic analysis and the inhibitory activities of isochlorogenic acids A and B were confirmed by whole-cell patch clamp recordings of human embryonic kidney 293 (HEK293) cells expressing human TRPV3. Molecular docking results revealed that these two compounds reside in the same active pocket of human TRPV3 channel protein with lower binding energy than the agonist 2-aminoethoxydiphenyl borate (2-APB). High-speed counter-current chromatography (HSCCC) coupled with a liquid-liquid extraction approach was successfully established for the separation of isochlorogenic acids A and B from the whole herbs of A. alpina. Ethyl acetate and n-hexane-ethyl acetate-water (3:3:4 and 1:5:4, v/v/v) were selected as liquid-liquid extraction solvent systems to remove high- and low-polarity impurities in the mixture. Sixty g of ethanol extract was refined by solvent partition to yield 1.7 g of the enriched fraction, of which 480 mg in turn obtained 52.5 mg of isochlorogenic acid B (purity 98.3%) and 37.6 mg isochlorogenic acid A (purity 96.2%) after HSCCC with n-hexane-ethyl acetate-water containing 1% acetic acid (1:4:8, v/v/v).

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Liquid—liquid extraction of membranes from calf brain using conventional and centrifugal counter-current distribution techniques

Journal of Chromatography B Biomedical Sciences and Applications ◽

10.1016/s0378-4347(00)84721-0 ◽

1984 ◽

Vol 311 ◽

pp. 277-289 ◽

Cited By ~ 15

Author(s):

Göte Johansson ◽

Hans-erik Åkerlund ◽

Björn Olde

Keyword(s):

Current Distribution ◽

Liquid Extraction ◽

Liquid Liquid Extraction ◽

Counter Current ◽

Counter Current Distribution

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Preparative Separation of Alkaloids from Stem of Euchresta tubulosa Dunn. by High-Speed Counter-Current Chromatography Using Stepwise Elution

Molecules ◽

10.3390/molecules24244602 ◽

2019 ◽

Vol 24 (24) ◽

pp. 4602 ◽

Cited By ~ 3

Author(s):

Weixin Li ◽

Huan Wang ◽

Aiwen Dong

Keyword(s):

High Speed ◽

Methylene Chloride ◽

Liquid Extraction ◽

Preparative Separation ◽

Target Compound ◽

Two Phase ◽

Chinese Herbal ◽

Liquid Liquid Extraction ◽

Counter Current ◽

First Time

Euchresta tubulosa Dunn. is a Chinese herbal medicine with biological activity, but there are few studies on its components at present. Alkaloids in the stem of Euchresta tubulosa Dunn. were isolated and purified by high-speed counter-current chromatography (HSCCC) using stepwise elution. First of all, liquid-liquid extraction (methylene chloride-methanol-water, 5:1:4, v/v) was used for the preliminary enrichment. According to the partition coefficient (K) of a target compound in a series of different two-phase solvents, the final result was that carbon tetrachloride-methylene chloride-methanol-water (2:3:3:2, v/v) (1) and methylene chloride-methanol-water (5:3:2, v/v) (2) were suitable for the HSCCC using stepwise elution. As a result, the purity was all higher than 93% and matrine (1), oxymatrine (2), N-formyl cytisine (3), and N-acetyl cytisine (4) can be eluted at one time by this mode. Cytisine-type alkaloids were isolated for the first time in this plant. Finally, the applicability of the mode was verified.

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