The present disclosure provides a method for detecting a display substrate and a device for detecting a display substrate. The method includes: exciting a threshold voltage of a driving transistor in each pixel driving circuit in the display substrate, so that the threshold voltage of the driving transistor with a shifted threshold voltage is further shifted; inputting a detection signal to each pixel driving circuit in the display substrate, where the detection signal is a signal enabling the pixel driving circuit to operate normally; and judging whether the display substrate is normal or not according to the voltage output by each pixel driving circuit.