Analysis of Angelica polysaccharides constituents by high-performance liquid chromatography

OBJECTIVE: To analyze the distribution of weight-average molecular weight (M̄r) of four Angelica polysaccharides isolated from Angelica sinensis (Oliv.) Diels: AP-I, AP-II, AP-III, AP-IV and determine their characters of electric-charge, in order to provide a practical analytical method for studying the constituent-activity relationship of polysaccharides. METHODS: Two kinds of high-performance liquid chromatography methods were employed, including high-performance gel filtration chromatography for M̄r determination and high performance anion exchange chromatography for electric-charge analysis. Improved phenol-sulfuric acid method combined with ELISA reader were used for the polysaccharide fractions determination, and Sever blue G dye-combined method for protein fractions as well. RESULTS: Each of the four Angelica polysaccharides, AP-I, AP-II, AP-III and AP-IV, was composed of 4~5 polysaccharide constituents with different M̄r distribution and characters of electric-charge; and there were small amount of different free or combined proteins in the four Angelica polysaccharides. CONCLUSION: The four Angelica polysaccharides, AP-I, AP-II, AP-III and AP-IV, are different from each other in M̄r and character of electric-charge; this method is practical and reliable for the study of constituent-activity relationship of polysaccharides.