Objective: To screen the active compounds targeted to beta2-adrenoceptor (β2-AR) in the water extract from Corydalis yanhusuo. Methods: β2-AR was immobilized onto macroporous silica gel by diazonium reaction to prepare the affinity extraction material. The material was characterized by plotting the adsorption isotherm of the specific drugs and the gel. Water extract of Corydalis yanhusuo was screened by the affinity extraction material. The bound and unbound components to β2-AR were seperated by centrafugation and the bound component was considered to be the bioactive compounds in Corydalis yanhusuo that target to β2-AR. The bioactive compounds were identified by high performance liquid chromatography (HPLC)-electrospray (ESI)-quadrupole time of flight tandem mass spectrometry (Q-TOF-MS). Results: The main bioactive compounds in Corydalis yanhusuo were alkaloids, including tetrahydrocolumbamine, corydaline, tetrahydroberberberine and tetrahydropalmatine. Conclusion: Compared with receptor chromatography, β2-AR afinity extraction method has the advantage in remaining the bioactivity of receptors. Because this method avoid the high pressure packing procedures in preparing the receptor chromatographic column. In addition, this method is fast, simple and has high efficiency in screening bioactive compounds in traditional Chinese medicine, thus provides a good reference for high throughput drug screening.