| Structure Identification: |
| J Sep Sci. 2011 Aug;34(15):1910-6. | | Counter-current chromatographic method for preparative scale isolation of picrosides from traditional Chinese medicine Picrorhiza scrophulariiflora.[Pubmed: 21721121] | Apocynin, androsin, together with picroside I, picrosideII and Picroside III from crude extracts of Picrorhiza scrophulariiflora were isolated by means of high-speed counter-current chromatography (CCC) combining elution-extrusion (EE) and cycling-elution (CE) approach. The EECCC took full advantages of the liquid nature of the stationary phase for a complete sample recovery and extended the solute hydrophobicity window, while CECCC showed its unique advantage in achieving effective separation of special compounds through preventing stationary phase loss.
METHODS AND RESULTS:
In the present work, the biphasic liquid system composed of n-hexane/ethyl acetate/methanol/water (1:2:1:2, v/v/v/v) was used for separation of apocynin and androsin, ethyl acetate/n-butanol/water/formic acid (4:1:5:0.005, v/v/v/v) for picroside I,picroside II and Picroside III.However, due to the extremely similar K values (K1 /K2 ≈1.2), picroside I and Picroside III were always eluted together by several biphasic solvent systems. In this case, the CECCC exhibited great superiority and baseline separated in the sixth cycle using ethyl acetate/water (1:1, v/v) as biphasic liquid system.
CONCLUSIONS:
Each fraction was analyzed by UPLC-UV and ESI-MS analysis, and identified by comparing with the data of reference substances. Compared with classical elution, the combination of EE and CE approach exhibits strong separation efficiency and great potential to be a high-throughput separation technique in the case of complex samples. |
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