The invention relates to a two-channel detection card for simultaneously detecting glibenclamide and gliclazide and a detection method thereof, belonging to the technical field of detection of western medicine illegally added into Chinese medicine. The surface of a detection card casing is provided with a detection window opening and a sampling hole, a test strip is arranged in the casing, a nitrocellulose membrane is attached to the middle of a bearing back panel, a water sucking membrane is attached to one end of the bearing back panel, a sample pad is attached to the other end of the bearing back panel, two colloidal gold membranes are clamped and attached between the nitrocellulose membrane and the sample pad, wherein the colloidal gold membranes are glass fiber membranes which respectively contain an anti-glibenclamide colloidal gold labeled monoclonal antibody and anti-gliclazide colloidal gold labeled monoclonal antibody, three print display bands are intervally arranged on the nitrocellulose membrane along the traverse direction, one is a detection band containing a glibenclamide protein conjugate, another one is a detection band containing a gliclazide protein conjugate, and the third one is a quality control band containing an anti-rabbit antibody or an anti-mouse antibody, the sample pad is arranged just opposite to the sampling hole, and the nitrocellulose membrane is arranged just opposite to the detection window opening. The detection card can be used for simultaneously detecting two western medicine components illegally added into a glucose-lowering Chinese medicine sample, has the advantages of detection cost saving, rapid detection, high sensitivity, accurate result, and convenience for use.