A signal gain determination circuit including a digital comparator, a digital controller and an arithmetic module, and a signal gain determination method are provided. A sensing integration circuit generates a first count during a first integration time according to a first sensing signal. The digital comparator compares the first count and a predetermined count to generate a comparison result. The digital controller generates a control signal for indicating a signal gain to a signal amplifier of the sensing integration circuit according to the comparison result. The signal amplifier adjusts the first sensing signal according to the signal gain to generate a second sensing signal, so that the sensing integration circuit generates a second count corresponding to the second sensing signal during a second integration time. The arithmetic module generates an output count corresponding to the first sensing signal according to the second count and the signal gain.