The pharmacokinetic in the aquatic was valued recently. It is advantageous to avoid blind with medicine in breed Penaeus chinensis. In this paper, studies on the pharmacokinetic parameter of chloromycetin in Penaeus chinensis. Chloromycetin is the stronger medicine in controling Penaeus disease. The 2/1000 contents of antibiotic chloromycetin was added into Penaeus chinensis feed producted with conventional method. The length of Penaeus sample was 9.5 cm. The test makes the Penaeus chinensis be feeded medicine only one time and get samples of Penaeus in different times then cooled immediately. Before analysis dissect, weigh, pre-treat properly and then determine contents in muscle, blood, intestines-stomach and hepato-pancreas. The method for pharmacokinetic study on antibiotic chloromycetin in Penaeus chinensis body was established by HPLC determination. The operating conditions were micro-Bondapak-C18 column, 30:70 (V/V) MeOH/H2O eluent and UV detector. The method is more rapid, accurate and sensitive. The correlation coefficient of the calibration curve is 0.9995 in the range of 1-50 mg/L.