This study aims to determine the effect of clove oil concentration and time of immersion in clove oil on the anesthesia, recovery time, and physiological responses in the payangka fish (Ophieleotris aporos). The experimental design was completely randomized with a 6 x 3 factorial arrangement. The first factor was a concentration of clove oil consisting of six levels i.e., 0.02; 0.03; 0.04; 0.05; 0.1, and 0.2 ml/L of water. The second factor was a time of immersion consisting of three levels i.e., 1; 3; and 6.5 hours. Results showed that the concentration of clove oil significantly (p<0.05) affected the anesthesia time and the recovery time after immersing. The length of immersing and concentration of clove oil also significantly (p<0.05) affected the concentration of glucose, but not significantly (p>0.05) affected the concentration of cholesterol. It showed that clove oil can be used as anesthetization for payangka fish in the appropriate dose. The optimal concentration of clove oil that can be used as anesthesia for the transportation of live payangka fish is 0.02 ml/L of water. Meanwhile, for handling activities on payangka fish, a clove oil concentration of 0.03 ml/L of water is recommended