Abstract: 【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.

Key words: mono-cellooligosaccharide, silica column chromatography, separation, FACE, ESI-TOF-MS