| This paper studied on the separation and purification of soybean phosphoplipids, soyasaponins, soy isoflavones from the by-products of soy molasses.Firstly, components of soy molasses and acid deposition were studied. Then the acid deposition was extracted with ethanol-hexane for the preparation of phospholipids concentrate; then the bleaching condition of phospholipids with hydrogen peroxide were studied; and powered phospholipids were collected through extraction of acetone. The optimal ratio of ethanol-hexane for the extraction was 1:1. The optimal conditions for the preparation of powered phospholipids were determined by one-factor contrast experiment. The optimal acetone extraction conditions were: acetone and sample ratio was 8:1, mixing time was 30min and extraction time was 3. The optimal bleaching conditions determined by one-factor contrast experiment were: the concentration of hydrogen peroxide was 4%, bleaching time was 60min, temperature was 50℃. The columns were used for purification of the powered phospholipids, the component which mainly contained phosphatidylcholine was collected and the product was analysised by HPLC.Secondly, the main components in the ethanol phase and the purification of soyasaponins were studied; then soyasaponins were purified by AB-8 macro absorption resin. According to the results of HPLC-MS, the main components of the ethanol pahse were 4 soy isoflavones: daidzin, genistin, glycitin, 6''-O-acetyl-genistin and 8 soyasaponins: soyasaponin Ba, Bb, Bc, A2, A4, Ea, Ab, Af. The optimum pH for separetion determined by static state experiment was pH3.0. The optimum conditions for separation determined by dynamic experiments were: room temperature, eluted by 50% ethanol for 3BV, 95% ethanol for 4BV, the 95% ethanol elution was collected. After concentration, extraction and crystallization, 98.46% soyasaponins were collected. The product was analysised by HPLC-MS, results showed that there were 4 soyasaponins in the product: soy saponin Bb, Bc, Ab, Af.At last, soy isoflavones were separated and purified from the 50% ethanol elutions through the treatment of calcium hydroxide, acid hydrolyzation and column separation. The optimum pH for the treatment of calcium hydroxide was pH9.0; the optimum conditions for hydrolyzation were: the concentration of hydrochloric acid was 2mol·L-1, temperature was 98℃, reaction time was 2h. The separation conditions for column were: H103 macro absorption resin, collect 75%-95% elutions. Then 73.85% products were collected. The products were analysised by HPLC and HPLC-MS, results showed that there were 3 isoflavones in the product: daidzein, glycitein and genistein. The content of daidzein was 45.03%.
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