Ciprofloxacin is the most commonly used fluoroquinolones for treating bacterial disease in olive flounder (Paralichthys olivaceus) farming, but its withdrawal time for industrial-scale farming has not been established. The depletion of ciprofloxacin was investigated in the olive flounder under field conditions. Fish were kept in an inland fish farm and fed a commercial diet containing 5 mg/kg of ciprofloxacin for 13 days. Seven fishes per sampling time were examined during and after the treatment. Ciprofloxacin and its major metabolite, enrofloxacin, were analyzed using HPLC with a fluorescence detector. The concentrations of ciprofloxacin and enrofloxacin in muscle increased during the medication period, and then decreased rapidly. The ciprofloxacin and enrofloxacin concentration in the olive flounder peaked on days 11 and 13, respectively, with maximum concentrations in muscle of 0.58 and 0.73 mg/kg. Residual ciprofloxacin and enrofloxacin were eliminated rapidly; 6 days after treatment, the respective levels in muscle were 0.04 and 0.10 mg/kg and neither was detected 15 days post treatment. The level of ciprofloxacin accumulation at the beginning of oral administration was variable according to the farming conditions, but the overall exhaustion time was similar. In conclusion, an adequate withdrawal period for enrofloxacin is 15 days in the case of oral ciprofloxacin administration.