Thermal comfort is one of the most important factors affecting human performance. In recent years, the Low Frequency to High Frequency Ratio (LF/HF) index, which is obtained from the frequency analysis of heart rate variability, has been proposed as an accurate method for evaluating thermal comfort in different conditions. In this study, using LF/HF, the effect of sweating amount on thermal comfort under the influence of clothing characteristics was investigated. After a moderately heavy physical activity in the indoor situation and without good ventilation, skin temperature, electrocardiogram signal and a qualitative assessment of their thermal comfort were recorded in participants divided into two groups: low sweat and high sweat. The results of qualitative measurement and frequency analysis of thermal comfort were then discovered to have a significant correlation. According to findings, the low sweat group had less thermal comfort than the high sweat group ( p < 0.05). The type of clothing was also found to have a significant effect on the thermal comfort sensation. Although the amount of sweating is an inherent feature of an individual, the feeling of thermal comfort can be improved ( p < 0.05) by optimizing clothing characteristics for each group without spending more energy on cooling.