Abstract: To establish a quality evaluation method for Astragali Radix using polysaccharide as quality control index, we established the Astragalus polysaccharide and monosaccharide sugar spectra, and combined with immunological activity test. High performance liquid chromatography (HPLC) was used to establish the specific chromatograms of Astragalus polysaccharides and monosaccharides. The data were analyzed by multivariate statistical analysis and cluster analysis using SIMCA software and SPSS software to distinguish Astragalus membranaceus var. mongholicus from different habitats or planting methods. The activity was evaluated by testing mouse peritoneal macrophage phagocytosis using neutral red. The results showed that the content of polysaccharides and the ability of enhancing phagocytic activity of macrophages from imitation wild Astragali Radix in Shanxi Hunyuan was higher than cultured Astragali Radix. The polysaccharides of Astragali Radix from Shanxi Hunyuan, Shanxi Wuzhai and Gansu Longxi have similar molecular weight distribution, but the peak area of each part has a significant difference in the percentage of the total peak area. The part of the polysaccharide of Shanxi Astragalus membranaceus with a molecular weight of about 10 kDa is higher than that of Gansu. Principal component analysis (PCA) shows that Astragali Radix from Shanxi and Gansu can be separated. All three are composed of five monosaccharides such as rhamnose, glucose, galactose, arabinose and galacturonic acid. However, the Astragalus polysaccharides (APS) in the three regions have different ratios of monosaccharide substances. The PCA display can distinguish three different Astragalus membranaceus var. mongholicus. This study used a combination of fingerprint of carbohydrates and the effects of APS on cellular immune function to provide a basis for quality evaluation and quality control of different habitats or planting methods.