A simple and sensitive high performance liquid chromatographic method for simultaneous determination of eight kinds of neutral monosaccharides, two kinds of uronic acid and one kinds of glycosamine by precolumn derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) has been developed. The polysaccharides hydrolyzed effects of hydrolysis time, temperature and trifluoroacetic acid (TFA) concentration were investigated. HPLC were established for analyzing the monosaccharide composition and the content of polysaccharide from Nostoc sphaeroides Kützing extractive. The results showed that the recoveries of nine kinds of monosaccharides, such as Man, GlcUA, Glc, Xyl, were 81.08%～102.1%. Compared with the chemical method. The established method is accurate, goodlineal relation，reproducible and specific, which could be used for the determination of qualitative and quantitative analysis in Nostoc sphaero-ides Kützing extractive.