High-quality digital sound sources with inaudible high-frequency components (above 20 kHz) have become available because of recent advances in information technology. Listening to such sounds has been shown to increase the α-band power of an electroencephalogram (EEG). The present study scrutinized the time course of this effect by recording EEG along with autonomic measures (skin conductance level and heart rate) and facial electromyograms (corrugator supercilii and zygomaticus major). Twenty university students (19-24 years old) listened to two types of a 200-s musical excerpt (J. S. Bach's French Suite No. 5) with or without inaudible high-frequency components using a double-blind method. They were asked to rate the sound quality and to judge which excerpt contained high-frequency components. High-α EEG power (10.5-13 Hz) was larger for the excerpt with high-frequency components than for the excerpt without them. This effect was statistically significant only in the last quarter of the period (150-200 s). Participants were not able to distinguish between the excerpts, which did not produce any discernible differences in subjective, autonomic, and facial muscle measures. This study shows that inaudible high-frequency components have an impact on human brain activity without conscious awareness. Unlike a standard test for sound quality, at least 150 s of exposure is required to examine this effect in future research.