Analysis of monosacchaaride compositions in polysaccharides by pre-column derivatization high performance liquid chromatography

A simple and sensitive pre-column derivatization high performance liquid chromatographic method (HPLC) for the analysis of monosacchoride compositions in polysaccharides is described. Six reducing monosaccharides as their 1-phenyl-3 - methyl-5-pyrazolone (PMP) derivatives were separated by reverse-phase HPLC using a developed gradient elution process, and monitored by ultraviolet detector at 250 nm. The broad reagent peak of PMP was separated very well from all the PMP-sugers, and good separation was achieved for derivatives Of mannose, rhamnose, glucuronic acid, glucose, galactose and xylose. The monosaccharide composition in polysaccharide was determined by a data analysis method which was developed to ensure accurate result and good reproducibility.