高速逆流色谱结合制备型高效液相色谱法分离制备大豆皂苷单体  

Large-Scale Isolation and Preparation of Soybean Saponin by High-Speed Countercurrent Chromatography Combined with Preparative HPLC

作　　者：黄玉艾[1] 严明霞[1] 赵大云[1] 

机构地区：[1]上海交通大学农业与生物学院,上海200240

出　　处：《食品科学》2013年第6期27-32,共6页Food Science

基　　金：国家自然科学基金面上项目(31071630)

摘　　要：为建立一种有效的同时分离制备多种大豆皂苷单体的方法,先采用高速逆流色谱分离去除大豆皂苷粗提物中的大豆异黄酮,然后对得到大豆总皂苷,优化制备型高效液相的流动相酸(乙酸或三氟乙酸)的含量和梯度洗脱程序,以制备高纯度的大豆皂苷单体。结果表明:流速10mL/min,流动相乙腈(含0.01%三氟乙酸)-水按梯度程序Ⅲ洗脱,出峰数量多、峰形好、分离度高。经高效液相分析与光谱特性、质谱和核磁共振鉴定了11个峰的大豆皂苷种类,包括5种A组、2种B组以及3种2,3-2H-2,5-二羟基-6-甲基-4H-吡喃-4-(DDMP)大豆皂苷。研究结果提供了从含量相对丰富的大豆胚芽中大量分离制备部分或完全乙酰化的A组以及非DDMP和完整DDMP存在的B组大豆皂苷的有效方法。The objective of this study is to isolate and purify soybean saponin from soybean germs by high-speed counter- current chromatography （HSCCC） combined with preparative HPLC. The isoflavonoids were removed from crude extract by HSCCC first. The pure soybean saponin was then obtained by preparative HPLC with acetic acid or trifluoroacetic acid （TFA） as mobile phase in a gradient elution mode. The optimal conditions for preparative HPLC were dual mobile phases composed of acetonitrile containing 0.01% （V/F） TFA and millipore water at a flow rate of 10 mL/min for gradient elution. Five compounds in group A and two compounds in group B and 3 kinds of 2,3-dihydro-2,5-dinydroxy-6-methyl-4H-pyran-4-one （DDMP）- conjugated soybean saponins with high purity were prepared and their structures were elucidated by mass spectrum （MS） and nuclear magnetic resonance （NMR）. Therefore, the developed method can be used to separate partially and fully acetylated soybean saponins as well as DDMP-conjugated and unconjugated soybean saponins in the original sample.

关 键 词：大豆皂苷单体 高速逆流色谱 制备型高效液相色谱 质谱 核磁共振 

分 类 号：TS207.3[轻工技术与工程—食品科学]