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 ?m) 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 ?l;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.