In order to investigate the shear behavior of composite beam with web opening under negative moment,simply supported composite beam under concentrated load were tested.The test results showed that the stiffness and ultimate capacity decreases significantly after web opening and the plane hypothesis is not satisfied any more.The failure mode is shear fracture in concrete slab near the opening area,and some stud connectors in opening area are pulled off.The ultimate load bearing capacity and deformation capacity can be effectively enhanced by increasing the thickness and reinforcement ratio of a concrete slab.This research provides an experimental basis for the EFA and the application of composite beams with web openings under negative moment.