制备型高速逆流色谱分离纯化香菇多糖

Separation and Purification of Lentinan by Preparative High Speed Counter Current Chromatography

利用高速逆流色谱仪,研究了双水相系统对香菇多糖的分离。溶剂系统为w(PEG1000):w(K2HPO4):w(KH2PO4):w(H2O)=0.5:1.25:1.25:7.0,在转速为500r/min,流速为1.5mL/min的条件下,成功分离了香菇多糖粗品(700mg),得到LenⅠ(95mg)、LenⅡ(45mg)两个组分。用SephadexG-100凝胶色谱柱检测纯度,结果显示:LenⅡ为单一峰,凝胶渗透色谱法测定;LenⅡ分子量为293kDa;经酸水解后液相色谱分析表明,其单糖组成为葡萄糖和甘露糖,摩尔比为2.7:1;红外光谱显示其具有多糖类的特征吸收峰。

High speed counter current chromatography（HSCCC） had been applied for the separation of lentinan. The HSCCC run was carried out with a two-phase solvent system composed of polyethylene glycol 1000 （ PEGI000 ） -dipotassium hydrogen phosphate-monopotassium phosphate- water（0.5 ： 1.25 ： 1.25 ： 7.0, m/m ） by eluting the lower aqueous phase at 1.5 mL/min at 500 r/min. Under the optimized conditions, One-step HSCCC procedure yielded 95 nag of Len Ⅰ and 45 nag of Len Ⅱ. By Sephadex G-100 column chromatography, Len Ⅱ was identified to be homogenous, and the molecular weight was 293 kDa according to the method of gelfiltration. The IR sepectrum of Len Ⅱ showed polysaccharide a character.