甘露糖酯的分离纯化及分析方法研究

本文对脂肪酶酶催化合成月桂酸甘露糖酯的分离纯化作了研究。确定了甘露糖酯的薄层层析(TLC)条件:反应液3μl点样,在正己烷/乙酸乙酯(1:1,V/V)中展开,用5%硫酸乙醇溶液喷雾,在120℃烘箱显色20min。将TLC条件应用到硅胶柱层析分离甘露糖酯,条件为:反应液5ml上硅胶层析柱(硅胶60～100目,柱12×600mm),流动相为正己烷/乙酸乙酯(1:1,V/V),流速为18ml/h,按1管/10min收集洗出液,并用TLC检测、收集产物。采用质谱(MS)方法鉴定了分离纯化的甘露糖酯产物,发现丙酮溶剂中脂肪酶催化合成产物为月桂酸甘露糖的单酯和两种二酯异构体。而用HPLC-MS分析方法证实乙腈溶剂中合成产物为月桂酸甘露糖的单酯、二酯和三酯。

Study of Purification and Analysis of Lauroyl Mannoses

The purification and analysis methods were studied for lipase-catalyzed synthesis of lauroyl mannoses. The qualitative analysis conditions for the thin layer chromatography (TLC) were confirmed: 3μl of the reaction mixture were applied to silica gel G plates, while hexane-ethyl acetate (1:1, V/V) was used as mobile phase. The reaction mixture was detected by spraying with 5% of sulfuric acid in ethanol and heated at 120℃ for 20min. Then, the conditions of TLC were applied to silica gel column chromatography for separating and purifying the lauroyl mannoses: 5ml of the reaction mixture are applied to silica gel column (60～100mesh) while hexane-ethyl acetate (1:1,V/V) is used as mobile phase. The flow rate is 18ml/h and the eluent of 1tube/10min is collected and detected with TLC. The purified products of the lauroyl mannoses synthesized in acetone are analyzed by MS, and the products are identified to be mono- and two kinds of dilauroyl mannoses. The products synthesized in acetonitrile are identified with HPLC-MS as mono-, di-, and trilauroyl mannose, respectively.