高速逆流色谱分离大豆皂苷和异黄酮

Separation of Soybean Isoflavones and Soyasaponins by High-Speed Countercurrent Chromatography

为建立一种同时分离大豆皂苷和异黄酮的制备方法,用高速逆流色谱,在正丁醇-乙酸-水溶剂体系下分离大豆异黄酮和皂苷,并通过调整乙酸含量得到最优的溶剂体系。结果表明,在该体系下可得到分离度很高的两个峰,第一个峰主要为大豆异黄酮,第二个峰为大豆皂苷。随着乙酸含量的增加,皂苷逐渐集中于第二个峰,大豆异黄酮和大豆皂苷被逐渐分开。正丁醇-5%乙酸-水,体系对大豆异黄酮和皂苷达到最好的分离效果。大豆粗提物进样量过大时,不利于两者的分离,在流动相流速为3 mL/min时,最佳的进样量为100 mg。

High-speed countercurrent chromatography（HSCCC） was applied to isolate soybean isoflavones and saponins from their crude extract under solvent system of n-butanol-acetic acid-water,and the concentration of acetic acid in the solvent system was optimized in order to achieve best separation.Results： With the HSCCC method isoflavones and soyasaponins are well separated and two distinct peaks appear,one of which is mainly isoflavones and another is soyasaponins in dominant.The two components are separated better as acetic acid concentration grows,and they are completely separated under acetic acid concentration of 5%（V/V）.A large sample loading is adverse to the separation,and the optimal sample loading is around 100mg at mobile phase flow rate of 3.0 mL/min.