Separation And Purification Of Natural Phospholipids

Natural phospholipids are recognized as bioactive substances with relatively valuable for pharmaceutical , because they exhibit many important physiological activities. They have been widely used in the field of nourishment,pharmaceutical and cosmetic industries.The high purity individual phospholipids have been demanded in above industries. Thus, the demand for high purity individual phospholipids enhances greatly. However, it is very difficult to obtain the high purity individual phospholipids. The phosphatidylcholine can be purified with simple column chromatography while the other phospholipids are separated by using HPLC. Owing to the elution mode to be used in HPLC,in which there are some inherent shortcomings,i.e.,low loading amount,high solvent consumption,low concentration of product,high production costs that cased high price of individual phospholipids production .How to reduce the cost of production is a vital problem that whether the high purity phospholipids could meet the demand of pharmaceutical or not.Aiming at the actuality of the utilization of the phospholipids, we used soybean oil degummed residues as raw material to prepare individual of soybean phospholipids by using column chromatography with simple eluent system and displacement chromatography for reducing the cost. The main contents studied are as follows: 1. Separation and purification of phosphatidylinositol from soybean oil degummed residues with solvent extraction and column chromatography has been studied. The effect of solvent composition in mobile phase and loading amount on separation efficiency were investigated basing on two indices of purity and recovery. Before separation, the raw material was pretreated to remove water and oil , and concentrated by removing ethanol-soluble lipids after ethanol (95% purity) extraction. The results shown that the optimum loading amount was 1.0 g/100 g silica when using silica as stationary phase and chloroform-methanol (1.8:1, v/v) as mobile phase under isocratic elution . The purity and recovery of phosphatidylinositol produced with above conditions was about 90%.While using diol column,the PI of high purity and recovery (>90%) can be obtained when using chloroform-methanol (2:1) as eluent, and loading 1.0 g sample/100 g packings.2. Separation of a tertiary mixture of PC , PE and PI by using displacement chromatography on aΦ12.5 mm×200 mm diol column has been studied for the first time. In the opinion of preparative separation,the effects of displacement concentration,flow-rate,loading amount on separation result is studied with purity and recovery as indices. When using dichloromethane-methanol(9:1,V/V) as mobile phase ,ethanolamine (7.5%,V/V) as displacer, setting flow-rate at 0.4 ml/min and loading 100 mg sample,the purity of PE(78.8%),PC(45.3%),PI(84%) can be obtained, and their recovery is 4 8 . 4 %,8 1 . 6 %,6 5 . 9 % .