AIM:The polysaccharides of Hedyotis difusa Willd of different sources were extracted and its content and composition of the polysaccharide was analyzed. METHODS: The microwave was employed for the aqueous extract of Hedyotis difusa Willd and the content of total sugar was determined by phenol-sulfur test,the monosaccharides composition was determined by HPLC with pre-column derivatization. RESULTS: The polysaccharide obtained from the samples of Guangdong Province had the highest content.The analytic data showed that the composition of monosaccharides in hydrolytic polysaccharides of Hedyotis difusa Willd were mannose,glucose,galactose,arabinose and rhamnose,the average content of polysaccharides was 9.82%. CONCLUSION: The HPLC method with pre-column derivatization is appropriate for the analysis of monosaccharide composition of Hedyotis difusa Willd polysaccharides and the method is simple,quick and accurate.