| In vitro: |
| Journal of Liquid Chromatography & Related Technologies, Volume 30, Number 4, March 2007, pp. 521-532(12) | | Preparative Isolation and Purification of Saponin and Flavone Glycoside Compounds from Clinopodium chinensis (Benth) O. Kuntze by High-Speed Countercurrent Chromatography[Reference: WebLink] | A preparative high-speed countercurrent chromatography (HSCCC) method for isolation and purification of didymin, nairutin, and Clinopodiside A from Clinopodium chinensis (Benth) O. Kuntze (Duanxueliu in Chinese) was successfully established.
METHODS AND RESULTS:
The separation was performed in two steps with two different kinds of solvent systems. In the first step, ethyl acetate-1-butanol-water (5:0.8:5, v/v) was used as the two-phase solvent system; nairutin was purified, and didymin and Clinopodiside A were eluted together. In the second step, ethyl acetate-methanol-water (5:1:5, v/v) was used as the two-phase solvent system; didymin and Clinopodiside A were separated and purified. After two-step separation, 15.2 mg of nairutin, 39.1 mg of Clinopodiside A, and 20.6 mg of didymin were obtained from 100 mg of crude extract with purities of 96.5%, 98.4%, and 99.1%, respectively, as determined by HPLC analysis. The chemical structures of the three components were confirmed by 1H NMR and 13C NMR. |
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