Abstract: An HPLC method was established for the determination of adenosine, γ-aminobutyric acid (GABA) and six flavonoids (calycosin-7-glucoside, ononin, calycosin, isoliquiritigenin, formononetin and medicarpin) in Radix Hedysari. The samples were extracted with methanol by refluxing for 4 h. The HPLCDAD was performed on a Diamonsil C₁₈ column (250 mm×4.6 mm, 5 μm) with acetonitrile-water as the mobile phase. The column temperature was at 40℃ and the flow rate was 1.0 mL·min^-1, while the temperature of drift tube was 110.5℃ and the nebulizing gas flow was 3.1 L·min^-1 for the ELSD system. The results showed all the eight components had good linear relationships (r²=0.992 8-1.000 0) in the range of the test concentration. The RSD of precision, stability and repeatability were less than 2%. The average recovery rates were 96.78%-103.45%, and RSD were 0.29%-1.61%. The index component contents of Radix Hedysari form 24 different origins were determined and used as variable factors in clustering analysis. The results were classified into 2 groups basically in accordance with the regional cluster. And the consequence was in consistent with the results of principal component analysis. This HPLC method is simple, shows good sensitive and accurate, and provides the experimental basis for multi-index control of Radix Hedysari. Clustering analysis for Radix Hedysari quality control has a certain reliability and objectivity.