制备型硅胶柱层析分离单一聚合度纤维寡糖及其测定

 【目的】探索实验室低成本、规模化制备单一聚合度纤维寡糖的方法，并得到聚合度分别为2—5纯度较高的单一组分纤维寡糖。【方法】采用制备型常压硅胶柱层析（ø22 mm×900 mm），选择干法填柱、干法上样对混酸降解法制备得到的纤维寡糖进行分离纯化。采用薄层色谱法（TLC）对分离过程进行监测，用荧光辅助糖电泳（FACE）对分离后的各单一组分进行定性检测，并用电喷雾-飞行时间质谱（ESI-TOF-MS）对各单一组分进行分子质量的确认。【结果】采用制备型常压硅胶柱层析法对于聚合度在2—5的纤维寡糖具有很好的分离效果。ESI-TOF-MS法检测本试验得到的4种单一聚合度纤维寡糖分子质量（钠离子峰）分别为365、527、689和851，分别对应纤维二糖、纤维三糖、纤维四糖和纤维五糖的分子质量。本试验一次上样300 mg纤维寡糖混合物，可分别获得28 mg纤维二糖、43 mg纤维三糖、59 mg纤维四糖和52 mg纤维五糖。荧光辅助糖电泳（FACE）能非常灵敏地定性检测纤维寡糖。【结论】制备型常压硅胶柱层析能较好地分离纤维寡糖，得到聚合度分别在2—5的单一组分纤维寡糖。

Separation and Detection of Mono-Cellooligosaccharide Purified by Preparative Silica Column Chromatography

【Objective】The aim of this paper was to explore a simple method to get sizeable amount of mono- cellooligosaccharide with degrees of polymerization（DP）two to DP five. 【Method】 The preparative silica column chromatography was selected to separate and purify the cello-oligosaccharides prepared from absorbent cotton. The separation process was monitored by TLC. FACE was used to test the separated single-component oligosaccharide and ESI-TOF-MS was used to detect their molecular weights. 【Result】 The experimental result showed that the preparative silica column chromatography had a better resolving ability for separating the cello-oligosaccharides with DP between two to five. From the ESI-TOF-MS spectrum, the molecular weights included one sodium ion of the four single-component cello-oligosaccharide were 365 for cellobiose, 527 for cellotriose, 689 for cellotetraose, and 851 for cellopentaose, respectively. Separated 300 mg cello-oligosaccharides once time, could get cellobiose 28 mg, cellotriose 43 mg, cellotetraose 59 mg, and cellopentaose 52 mg, respectively. FACE method was very sensitive to the qualitative analysis of cello-oligosaccharides. 【Conclusion】 The preparative silica column chromatography is a better separation and purification method for preparing mono-cellooligosaccharide with DP two to five.