This note describes a simple and rapid method to separate specific soyasaponins, including the major and multi-functional bioactive species soyasaponin Bb, from the whole soybean flour. The method is based on the difference in solubility of the soyasaponins and other components in methanol, aqueous borax solution, and 1-octanol. First, the whole soybean flour was mixed with methanol to extract hydrophobic components, and the methanol solution was concentrated. Second, the methanol solution was mixed with a larger volume of aqueous borax solution, and the supernatant was acidified to obtain a precipitate. The precipitate was found to be containing mainly four group B soyasaponins. Two of those are non-conjugated molecules, soyasaponins Bb and Bc, and the other two are 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one conjugated ones, soyasaponins βg and βa. All of them have two cis-diol groups in their structure. Therefore, although they are fundamentally insoluble in water, they became soluble in the aqueous medium in the form of divalent anionic species by the borate ester formation, and that they were re-precipitated by the hydrolysis. Furthermore, the non-conjugated group B soyasaponins could be separated by washing with 1-octanol to remove highly hydrophobic soyasaponins βg and βa. The solubility-based technique would be useful as pretreatment in the purification of these group B soyasaponins.
Keywords: Separation; soyasaponin Bb; soyasaponin Bc; soyasaponin βa; soyasaponin βg; whole soybean flour.