Separation of soybean isoflavone glucosides from Semen Sojae Praeparatum by preparative reversed-phase high performance liquid chromatography / 第二军医大学学报

ABSTRACT

Objective: To search for a rapid and efficient method for the isolation of 3 kinds of isoflavone glucosides from the ethanol extract of Semen Sojae Praeparatum. Methods: The crude isoflavones were extracted by 75% aqueous ethanol after removing the oil by Soxhlet extraction. Then 1300 macroporous resin, water and different concentrations of aqueous ethanol (10%, 20%, 30%, 40%, 50%, 70%, and 95%) were used to separate and elute the crude isoflavones. The fraction eluted by 40% aqueous ethanol was subjected to preparative HPLC analysis. The chromatographic conditions A YWG C18 (10.0 mm × 200 mm, i. d. 10 μm) column was used; the mobile phase consisted of acetonitrile-water-acetic acid at volume ratio of 25 75 1 (V/V/V) and a flow rate of 3.0 ml/min; the injection volume was 750 μl; and the detection wave length was set at 260 nm. Results: Daidzin, glycitin and genistin were rapidly separated with the purities over 99% as determined by external standard HPLC. Conclusion: This technique is simple and suitable for the isolation of daidzin, glycitin and genistin from Semen Sojae Praeparatum.