The gap‐startle paradigm has been used as a behavioral method for tinnitus screening in animal studies. This study aimed to investigate gap prepulse inhibition (GPI) of the auditory late response (ALR) as the objective response of the gap‐intense sound paradigm in humans. ALRs were recorded in response to gap‐intense and no‐gap‐intense sound stimuli in 27 healthy subjects. The amplitudes of the baseline‐to‐peak (N1, P2, and N2) and the peak‐to‐peak (N1P2 and P2N2) were compared between two averaged ALRs. The variations in the inhibition ratios of N1P2 and P2N2 during the experiment were analyzed by increasing stimuli repetitions. The effect of stimulus parameter adjustments on GPI ratios was evaluated. No‐gap‐intense sound stimuli elicited greater peak amplitudes than gap‐intense sound stimuli, and significant differences were found across all peaks. The overall mean inhibition ratios were significantly lower than 1.0, where the value 1.0 indicates that there were no differences between gap‐intense and no‐gap‐intense sound responses. The initial decline in GPI ratios was shown in N1P2 and P2N2 complexes, and this reduction was nearly complete after 100 stimulus repetitions. Significant effects of gap length and interstimulus interval on GPI ratios were observed. We found significant inhibition of ALR peak amplitudes in performing the gap‐intense sound paradigm in healthy subjects. The N1P2 complex represented GPI well in terms of suppression degree and test‐retest reliability. Our findings offer practical information for the comparative study of healthy subjects and tinnitus patients using the gap‐intense sound paradigm with the ALR.