The binaural masking level difference (BMLD) is a psychoacoustic method to determine binaural interaction and central auditory processes. The BMLD is the difference in hearing thresholds in homophasic and antiphasic conditions. The duration, phase and frequency of the stimuli can affect the BMLD. The main aim of the study is to evaluate the BMLD for stimuli of different durations and frequencies which could also be used in future electrophysiological studies. To this end we developed a GUI to present different frequency signals of variable duration and determine the BMLD. Three different durations and five different frequencies are explored. The results of the study confirm that the hearing threshold for the antiphasic condition is lower than the hearing threshold for the homophasic condition and that differences are significant for signals of 18ms and 48ms duration. Future objective binaural processing studies will be based on 18ms and 48ms stimuli with the same frequencies as used in the current study.