School of Chinese Medicine; Department of Chemistry
Using two-phase solvent systems for sample pretreatment increases yield of counter-current chromatography: Anemarrhena asphodeloides saponins, a case study
This paper reports a way to increase the production yield of counter-current chromatography by using two-phase solvent systems in sample pretreatment. This strategy succeeds in the case of anti-EV71 saponin preparation from Anemarrhena asphodeloides. Briefly, 15 g of water extract was refined by solvent partition with ethyl acetate/1-butanol/water (1:4:5, v/v) to yield 4.17 g of the upper part, which in turns yielded 635 mg of timosaponin B-II (purity 96.5%) and 672 mg of anemarsaponin B (97.8%) after gradient CCC separation with 1-butanol/water (containing 0.01 mol/L CuSO4 in water, 1:1, v/v) and ethyl acetate/1-butanol/water (3:17:20, v/v/v). In a similar way, the ethanol extract (8 g) was partitioned between the diphase solvent system of ethyl acetate/1-butanol/water (4:1:5) to get 2.16 g of refined fraction and yielded 358 mg of timosaponin A-III (96.4%) by CCC elution using ethyl acetate-methanol-water (4:1:5, v/v/v). To remove mangiferin, the salt CuSO4 was added; it formed a complex with mangiferin, thereby enlarging the separation coefficient between mangiferin and timosaponin B-II.
Anemarrhena asphodeloides, Anemarsaponin B, Counter-current chromatography, Timosaponin A-III, Timosaponin B-II
Source Publication Title
Separation and Purification Technology
© 2014 Elsevier B.V. All rights reserved.
This work was supported by the National Natural Science Foundation of China (Grant No. 81303289) and the Hong Kong Baptist University (FRG2/12-13/006, FRG2/13-14/026, and RC-start up grant).
Link to Publisher's Edition
Liu, Meng-Shun, Siu-Leung Chau, Dik-Lung Ma, Chung-Hang Leung, Zhao-Xiang Bian, Ai-Ping Lu, Hong-Xi Xu, and Quan-Bin Han. "Using two-phase solvent systems for sample pretreatment increases yield of counter-current chromatography: Anemarrhena asphodeloides saponins, a case study." Separation and Purification Technology 138 (2014): 138-143.