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As the complexity of the composition and structure of polysaccharide, there is still lack of analysis methods and quality standards which could effectively characterize and identify the polysaccharide from Ganoderma lucidum with physiological activity.In our study, two methods of partial acid hydrolysis coupled PMP pre-column derivatization reverse phase high performance liquid chromatography (RP-HPLC) and acid-enzyme hydrolysis coupled high-performance anion-exchange chromatography (HPAEC) respectively were developed for detect and characterize the compositive and structural difference of polysaccharides extracted from the Ganoderma lucidum spores of different sources, and establish two kind of chromatographic fingerprints of the polysaccharides.Furthermore, we investigated the promoting effect of the polysaccharide from Ganoderma lucidum spores on lymphocyte proliferation by means of MTT experiments, which was applied to research the relationship of RP-HPLC and HPAEC profile-immune activity and gave effective quality evaluation of Ganoderma lucidum spore and its products on the market.