大豆卵磷脂的分析与精制方法研究

【作者】 李艳；

【导师】 杨亦文；

【作者基本信息】 浙江大学 ， 应用化学， 2004， 硕士

【摘要】 本文在对大豆卵磷脂分析方法和提取方法进行综述的前提下，研究了大豆卵磷脂的分析和提纯技术。 经实验研究，确定了大豆磷脂的高效液相色谱—示差折光检测方法（HPLC—RI法），分析条件如下：色谱柱为Waters Spherisorb^? S5W（5μm Silica）4.6×250mm硅胶柱；流动相为正己烷：异丙醇：水（1:4:1，v／v／v）；流速：0.8ml／min；柱温：35℃；示差折光检测器温度：35℃。用大豆PC标样采取外标法对产品进行定量。此条件下PC出峰时间为11min左右，各种磷脂成分可以较好的分离。 通过三种粗提方法的比较，确定了大豆水化油脚的粗提方法为：分别用10倍体积的工业丙酮一次性处理和10倍体积的95％工业乙醇分三次处理，搅拌，抽滤，最后将乙醇提取液合并，蒸干，即可得到PC含量在40～50％大豆磷脂粗提产物，PC收率在65％左右（大豆水化油脚PC含量按15％计算）。 另外还研究了大豆卵磷脂的精制方法：先将粗提产物经过硅胶柱层析，除去PE、LPC和SM等杂质，然后再经过氧化铝柱层析，除去杂质磷脂PI。硅胶柱层析实验条件如下：层析介质为80～120目粗孔Ⅱ号硅胶；洗脱液为石油醚（60～90℃）：乙醇：水=1：1：0.175（V／V）；最佳上样量为0.037g粗提产物／ml床层体积；上样浓度：0.275g粗提产物／ml洗脱液。氧化铝柱层析洗脱液为95％工业乙醇。 大豆磷脂粗提产物通过这种精制方法，可以得到PC纯度为94.51％的产品，柱层析阶段收率为90.88％。更多还原

【Abstract】 In this text, we studied the separation, purification and determination technology of high purity soybean phosphatidylcholine.A simple rapid and accurate quantitative method was established for the determination of lecithin by HPLC-RI in this research. Separation was obtained by using Waters Spherisorb S5W (5μm Silica) 4.6 X 250 mm silica column and mobile phase of n-hexane:2-propanol:water(l:4:l,v/v/v) with flow rate of 0.8ml/min. Under this condition, the retention time of PC was about 11 minutes. This method produced data which correlated well with data from the TLC-P method, and this data was highly accurate and exhibited satisfactory reproducibility.The soy lecithin with 40-50% PC was obtained from soybean raw material by using acetone and 95% ethanol, the yield of PC in this process was about 65%.High purity soybean phosphatidylcholine can be obtained in this method: first PE LPC SM, etc was separated from PC by silica column chromatography, then the product of silica column chromatography was treated by alumina column chromatogr-aphy, PI can be removed and product with 94.51% PC can be obtained. The yield of this process was 90.88%.The mobile phase of silica column chromatography: mixed solvent of petroleum ether (60~90℃), ethanol and water(l:l:0.175); the mobile phase of alumina column chromatography :95% ethanol.更多还原