Oxytetracycline (OTC) has been widely used in aquaculture field as a therapeutic and prophylactic agent because of its broad-spectrum activity against gram-positive and negative bacteria. Residual oxytetracycline (OTC) was studied after spray treatment of cultured abalone, Haliotis discus hannai. Muscle concentration of OTC was determined after spray treatment ( 4,000, 7,000, 10,000 ppm) in the abalone. Muscle samples were taken at 0, 1, 3, 7, 14, 21 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, 330 and 360 day post-dose. OTC analyses were carried out by high performance liquid chromatography (HPLC). In 4000, 7000 and 1000 ppm treatment for small size abalones, OTC levels at 1 day post-dose, have been dramatically decreased by 8.34, 3.35 and 4.47 ug/g, respectively. For medium size abalones, concentration were measured as 7.58, 15.62 and 7.8 ug/g, respectively. Those of large size abalones also were observed as 11.31, 12.38 and 15.28 ug/g, respectively, at 1 day post-dose. No significant differences in residual OTC depletion in muscles were observed between the size of abalones. More than 0.2 mg/kg of OTC was detected in muscle tissues and the residues were found over 60 days after treatment. It is expected that these results would contribute to improve recommended withdrawl periods of OTC for a safer seafood supply.