中国食品学报2013，Vol.13Issue(1)：165-171,7.
Sephadex LH-20凝胶层析结合液相制备色谱分离纯化大豆皂苷单体
Seperation and Purification of Soyasaponin Individuals by Sephadex LH-20 Column and Preparative Liquid Chromatography
摘要
Abstract
Crude soyasaponins were extracted from defatted soybean germs with 70% ethanol aqueous solution. Gel filtration chromatography was employed to separate soyasaponins and isoflavones using Sephadex LH-20 as matrix and methanol as solvent. High purity of soysaponin-riched products were collected. The flow rate was 3 mL/min, and sample loading was 60mg of crude soysaponins in this method. A preparative reversed phase liquid chromatography was applied to isolate soyasaponin individuals. The mobile phase, elute program, and injection amount were optimized. As a result, the optimum operation conditions were as follow:a solvent system consisting of water(A) and acetonitrile with 0.01% tri-fluoroacetic acid (B) was used; the flow rate of mobile phase was 10 mL/min; and the injection amount was 70 mg of crude soysaponins dissolved in 0.5 mL methanol. UPLC-MS was used to elucidate the structures of prepared soyasaponin components. Nine soyasaponin individuals, ac3-Ab, Aa, Ab, Ae, Af, Bb, Be, Ba, Bb' with purity above 90% were obtained. DDMP-conjugated groups, αg, βg, γg, which are very labile even in mild conditions, were also collected but with low purity. The method described here can be used to separate partially and fully acetylated group A soyasaponins as well as DDMP-conjugated and unconjugated group B soyasaponins occuring in the original sample.关键词
Sephadex LH-20/制备型高效液相色谱/UPLC-MS/大豆皂苷单体Key words
Sephadex LH-20/ preparative HPLC/ UPLC-MS/ soyasaponin individuals引用本文复制引用
严明霞,黄玉艾,赵大云..Sephadex LH-20凝胶层析结合液相制备色谱分离纯化大豆皂苷单体[J].中国食品学报,2013,13(1):165-171,7.基金项目
国家自然科学基金(31071630/C200303) （31071630/C200303）
