In order to obtain the fundamental data for the marine ranch, this paper was carried out to investigate cortisol variation in blood according to adapted time in the measurement of auditory threshold. The groups were adapted at the experimental tanks for 0.5, 1, 3 and 6 hours, respectively. At the results, cortisol concentration had a deep connection with adapted time. In particular, cortisol concentration in the experimental group of 1 hour was significantly increased, whereas others showed no difference comparing with the control group. When classical conditioning method with the sound coupled with a delayed electric shock was given after the adapted time of 1 hour and 6 hours, the rate of the conditioning completion for 6 hours was higher than that of 1 hour. Therefore, this suggests that the sufficient adapted time was required for the accurate auditory threshold.