硅胶柱层析法提纯大豆磷脂酰胆碱的研究

Purification of Soybean Phosphatidylcholine by Silica Gel Column Chromatography

探讨了以大豆浓缩磷脂为原料，应用柱层析法提纯大豆磷脂酰胆碱（PC）的工艺过程。用TLC法考察了水、异丙醇与正己烷的体积配比，确定流动相适宜配比为正己烷：异丙醇：水（1：1：0．175，v／v／v）。通过对固定相、流动相和上载量的考察，得到了柱层析最佳工艺条件：在空塔流速为0．382cm·min^-1，进样浓度为0.25g浓缩磷脂·（mE流动相）^-1的室温条件下，固定相选用100-200目粗孔2号硅胶，流动相为正己烷：异丙醇：水（1：1：0．16，v／v／v），最适上载量为0.086gPC·（g硅胶）^-1。经高效液相色谱法定量分析，产品中磷脂酰胆碱纯度高达85％。

The process of purification of phosphatidylcholine （PC） from soybean concentrated phospholipids （SCP） was studied. The method selected was based on the normal-phase chromatography with silica gel as stationary phase and a ternary elution with n-hexane, isopropanol and water as mobile phase, Firstly, the suitable ratio of n-hexane, isopropanol and water in the mobile phase was investigated preliminarily by thin-layer chromatography （TLC）, The results show that the suitable ratio of mobile phase is n-hexane： isopropanol： water =1：1：0.175, by volume. Secondly, the optimal operation conditions of column chromatography separation were obtained by the investigation of the stationary phase used, mobile phase composition and column loading ability The results show that at the sample concentration of 0.25 gSCP·（mL mobile phase ）^-1 and the flow rate of 0.382 cm·min^-1, the 100-200 mesh silica gel can be chosen as the stationary phase, the ternary mixture of n-hexane：isopropanol： water = 1：1：0.16 （by volume） is the suitable mobile phase and the optimal loading is 0,086 gPC.（g silica gel ）^-1 Using above mentioned optimal operation conditions, the product purity of the column chromatography separation at room temperature was analyzed by HPLC （High-Performance Liquid Chromatography）, and the result shows that the purity of PC obtained can reach 85%.