The purpose of this paper is to investigate the autonomic nervous system activity in various ambient temperature situation. To evaluate autonomic function, we use the time domain analysis of heart rate variability. Electrocardiogram was recorded to derive heart rate variability in 3 different temperature room which temperature is controlled in 18℃(low), 25℃ (mid) and 38℃(high), respectively. Totally 22 subjects were participated in the experiment. The result shows that the time-domain variables such as AVNN, SDNN, SDSD, RMSSD, NN50, pNN50, NN20 and pNN20 show the significant difference between low and high temperature (p<0.01). However, these variables has no significance (p>0.05) between mid and high except on AVNN, RMSSD and pNN20. AVNN, RMSSD shows the highest significance (p<0.001) according to the various temperature environment.