In the present study, an efficient method based on ligand fishing and high-speed counter-current chromatography (HSCCC) was established to screen, enrich and separate the active components with the a-amylase inhibitory activity from a traditional dish Tornio sinensis. The active components were screened from T. sinensis by ligand fishing using the magnetic immobilized alpha-amylase prepared through solvothermal and crosslinking methods. HSCCC was used to separate the target compound according to the K value. As a result, a potential active compound 1.2,3,4,6-penta-O-galloyl-beta-D-glucose and a non-target compound quercetin-3-O-alpha-L-rhamnopyranoside were separated and identified. In-vitro experiments indicated that 1 2,3,4,6-penta-O-galloyl-beta-D-glucose had the activity against alpha-amylase and the IC50 value was 93.49 +/- 0.80 mu g/mL which was higher than that of the non-target compound. The result further confirmed the molecular fishing effect of magnetic immobilized alpha-amylase. The present study can not only find and separate the hypoglycemic substances in T. sinensis quickly and effectively, but also can provide a new approach for the study of natural active components.