Separation And Analysis Of Agaro-oligosaccharides By High Performance Capillary Electrophoresis

Agar is a kind of water soluble polysaccharide extracted from red algae like Gracilaria, it is composed of agarose and sulfuric agar. Agarose is a linear chain composed of alternately arranged 3-O-linked-β-D-galacto- pyranose and 4-O-linked 3,6-anhydro-α-L-galactopyranose residues.β- Agarase hydrolyzes ?-1,4 linkages of agarose yielding neoagaro–oligo saccharides, while the acid andα-Agarase hydrolyzesα-1,3 linkages yielding agaro-oligosaccharides. Research shows that neoagaro-oligosaccharides have the function of digestive enzyme tolerance and suppressing the growth of bacteria. Agaro-oligosacchaides can restrain the cancer cells and cure inflammatory diseases.In this paper agarose is degraded both by ?- Agarase and hydrochloric acid, the degradation products are separately neoagaro-oligosaccharides and agaro-oligosaccharides. The oligosaccharides are derivatized by 1-phenyl-3- methyl-5-pyrazolone(PMP) with high ultraviolet absorption in the pre- column derivatization and detected by CZE-UV. The optimization studies of derivatization, buffer concentration, buffer pH value and working voltage have been performed with respect to labeling and separation conditions of oligosaccharides. The result shows that the neoagaro-oligosaccharides reach the best separation in the sodium tetraborate decahydrate buffer of pH 9.0, 20 mmol/L at 25℃under the working voltage of 30 kV, while the agaro-oligo saccharides separate best in the sodium tetraborate decahydrate buffer of pH 10.0,30 mmol/L under the same temperature and voltage.The HPCE method is firstly used in the separation and analysis of neoagaro-oligosaccharides and agaro-oligosaccharides in this paper. It can also be used in the research of other oligosaccharides, and enlarged the analysis methods on oligosaccharides.