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Using air conditioner has been increased in home or office buildings in summer. Also various problems related to air conditioning such as disease induction happened by using air conditioner excessively and operating long. Active operation control is needed for occupant's health when air conditioner operates long. We should think ahead to acquire thermal comfort of occupants which represents psychological and physiological reaction for this operation. Research has been progressed to observe activity of autonomic nervous system by trying to quantitate change of thermal comfort. In this study, questions of the subject and change of body's autonomic nervous system were chosen to evaluate thermal comfort during operation of air conditioner for a long time. Electrocardiogram and questions of the subject which is the progress of changing TSV and CSV by occupants indoor were measured when room air conditioner is operated for a long time, and an air-conditioned adaptability of human body was evaluated by acquiring the change rate of autonomic nervous system through analyzing HRV. As a result of the evaluation, change rate of body's autonomic nervous system corresponded to votes of the subject's question generally, but was distinguished from analysis result of warm-cold sensation in a low temperature area.