Zhang Weinong, He Haibo, Feng Yuqi, Da Shilu
College of Chemical and Molecular Science, Wuhan University, 430072, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Dec 25;798(2):323-31. doi: 10.1016/j.jchromb.2003.10.005.
Natural phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were separated and purified from soybean degummed oil residues in this work. Crude PC and PE were first separated from degummed oil residues by extraction with 95% ethanol, and then the crude PC and PE were used as raw materials to prepare high purity PC and PE by using column chromatography of silica gel (100-200 mesh) with different eluents and elution modes. The high purity PC (content > 90%) was obtained from the crude PC by using isocratic elution with methanol as eluent. Compared with the methods reported by using isocratic elution with mixed solvents as eluent or gradient elution, the procedure proposed exhibits low cost and industry potentialities because of some advantages, such as operation simplicity, cheap equipment and solvent to be recovered easily. The purity of the PE product prepared from the crude PE was more than 75%. The gradient elution was preferable to isocratic elution for reducing the elution time and eluent consumption when to prepare PE from the crude PE. The effects of loading amount and the flow-rate on separation efficiency were also investigated. For obtaining high separation efficiency, the loading amount should be less than 2.0 g crude PC or PE/100 g silica gel, and the flow-rate should be controlled under 4 ml/min for crude PC and 3 ml/min for crude PE, respectively.
在本研究中，从大豆脱胶油残渣中分离并纯化了天然磷脂酰胆碱（PC）和磷脂酰乙醇胺（PE）。首先通过用95%乙醇萃取从脱胶油残渣中分离出粗PC和PE，然后以粗PC和PE为原料，采用硅胶柱色谱（100 - 200目），使用不同的洗脱剂和洗脱模式制备高纯度的PC和PE。以甲醇为洗脱剂等度洗脱从粗PC中获得了高纯度PC（含量>90%）。与使用混合溶剂等度洗脱或梯度洗脱报道的方法相比，所提出的方法具有操作简单、设备便宜、溶剂易于回收等优点，因而成本低且具有工业应用潜力。从粗PE制备的PE产品纯度超过75%。从粗PE制备PE时，梯度洗脱比等度洗脱更有利于减少洗脱时间和洗脱剂消耗。还研究了上样量和流速对分离效率的影响。为了获得高分离效率，上样量应小于2.0 g粗PC或PE/100 g硅胶，粗PC的流速应控制在4 ml/min以下，粗PE的流速应控制在3 ml/min以下。
