二十二碳六烯酸海藻糖酯的分离纯化及鉴定

Separation,purification and identification of docosahexaenoyl trehalose

在叔丁醇中以二十二碳六烯酸(DHA)乙酯和海藻糖为原料,通过固定化脂肪酶催化合成DHA海藻糖酯,并对其分离纯化方法进行研究。确定DHA海藻糖酯的薄层层析(TLC)条件:展开剂为乙酸乙酯/甲醇/水(8.5/1/0.5,v/v/v),碘蒸气显色10min;硅胶柱层析条件:正己烷/异丙醇/甲醇(5/4/1和4/4/2,v/v/v)梯度洗脱,流速1.3mL/min,1管/7min收集洗出液;用半制备高效液相色谱(HPLC)进一步纯化单酯收集液,经分析型HPLC检测纯度后用核磁共振(NMR)方法进行结构鉴定,确定为DHA藻糖单酯。

Docosahexaenoyl trehalose（DHA） was synthesized from docosahexaenoic acid ethyl ester and trehalose in tert-butanol using immobilized lipase as biocatalyst.The purification and analysis methods were investigated.The mobile phase for the thin layer chromatography（TLC） was ethyl acetate/methanol/water（8.5/1/0.5, v/v/v） and iodine was used for development.The silica gel column chromatography conditions were as follows： gradient mobile phases of hexane-isopropanol-methanol 5：4：1 （v/v/v） and 4：4：2（v/v/v） were used in turn.The flow rate was 1.3mL/min and the eluent of 1tube/7 min was collected.The eluent of mono-docosahexaenoyl trehalose was purified with semi-preparative high performance liquid chromatography HPLC further and then was identified by nuclear magnetic resonance（NMR）.