Based on the finite element analysis software OpenSEES, substructure pseudo-dynamic test (PDT) method for frame structures was investigated. Substructure PDTs of a one-story one-bay steel frame was conducted, in which three different kinds of boundary conditions were simulated. The correctness of the PDT method was demonstrated by the agreement between the test results with complete boundary condition and time history analysis results. For one-story, five-story, and eight-story four-bay reinforced concrete frames, substructure PDTs were also conducted with three different kinds of boundary conditions. The considerable discrepancy between the test and simulation results occurs in hysteresis curve of substructure columns when only horizontal displacement boundary condition was simulated. The simulation accuracy of the hysteresis curve of substructure columns can be improved when both the horizontal displacement and rotation boundary conditions are considered. The base shear was much less affected by using simplified boundary condition, while its effect on first story drift can be ignored. The results can be used for the design of substructure pseudo-dynamic tests.