磷脂在硅胶柱上穿透特性的实验研究

Experimental Studies on Breakthrough Characteristics of Phospholipids on Silica-gel Column

进行了不同流动相组成、进样浓度、流速、温度等对硅胶柱上磷脂穿透曲线影响的实验研究。结果表明在硅胶柱上卵磷脂(PC)比脑磷脂(PE)具有更强的结合力,因而在流出曲线中脑磷脂先于卵磷脂出峰;由于卵磷脂和脑磷脂的竞争吸附,使脑磷脂在流动相中甲醇含量较低时出现很高的过顶峰;同时,在流动相中甲醇含量较高时脑磷脂的流出曲线呈现双平台型,即显现虚拟二组分特性。以脑磷脂为目标产物,通过对磷脂处理量和脑磷脂理论产量的比较,得到了较佳的工艺条件:进样浓度为100mg·mL-1,流动相为纯氯仿,流动相流速为0.4mL·min-1,此时PE的理论产率可达0.054g?g(硅胶)-1。实验结果能为硅胶柱层析分离纯化磷脂提供重要的基础数据。

Effects of mobile phase composition, feed concentration, flow rate and temperature on the breakthrough curves of phospholipids on silica-gel column were experimentally studied. The results show that phosphatidyl ethanolamine （PE） comes out ahead of phosphatidyl choline （PC）. Higher overshoot peaks of the breakthrough curves of PE were found at lower content of methanol in the mobile phase, which may be attributed to the competitive adsorptions of PC and PE. In addition, the breakthrough curves of PE at higher content of methanol in the mobile phase exhibits double-plateau type, which indicates that PE could be treated as pseudo binary components. By comparing the input amount of crude phospholipids with the theoretical output of PE at the breakthrough point of PC, optimal operation conditions for purification of PE were obtained, i.e., using pure chloroform as mobile phase, flow rate at 0.4 mL·min^-1 and feed concentration of 100 mg·min^-1. The theoretical yield of PE could reach 0.054 g.g（silica-gel）^-1 by using this condition.