An improved carbohydrate derivatization method employing 1-phenyl-3-methyl-5-pyrazolone(PMP) was developed in this study.Liquid ammonia was used instead of sodium hydroxide aqueous solution to provide the base medium for the derivatization reaction.Derivatization reaction and HPLC separation conditions were optimized,including reaction time,reaction temperature and PMP consumption.The derivatives of five reducing monosaccharides were well separated by HPLC. Because we employed ammonia instead of sodium hydro-xide,the excessive volatile ammonia can be removed by vacumm drying,these derivatives could be directly analyzed by MALDI-TOF-MS without desalting procedure,possessing potential application for monosaccharide and oligosaccharide separation and subsequent MS analysis.This method not only increased the MS detection sensitivity,but also decreased sample wastes during purification. We successfully applied this method to mo-nosaccharide composition analysis of polysaccharides SPPA-1,SPPB-1 and SPPC-1 purified from Spirulina(Platensis).
