In this study, we analyzed two types of echolocation calls used by the parti-coloured bat, Vespertilio sinensis. Bats were captured in the Naejangsan National Park in October 2013. Call sounds of hand-released bats were recorded at the location of capture within the National Park. We analyzed pulse duration (PD), pulse interval (PI), peak frequency (PF), maximum frequency (FMAX), minimum frequency (FMIN), and bandwidth (BW). V. sinensis emitted the different types of the echolocation calls depending on the surrounding environment. Frequency modulated-constant frequency (FM-CF) signal of audible range was emitted when they flew in the uncluttered space over the canopy. However, when flying in the cluttered space below the canopy, they only emitted FM signal. FM-CF signal is in the audible range (e.g., low frequency), and FM signal has a harmonic broadband frequency range of two. There were significant differences in PD, PI, PF, FMAX, FMIN, and BW between the calls emitted over and below the canopy. Considering the functional characteristics of FM and CF signals, we conclude that the foraging activity of V. sinensis was observed below the canopy, and recommend the use of FM signal and broadband as echolocation signals.