The distribution and depletion of clenbuterol hdrochloride in ovine eyes,heart,lung,kidney and spleen of sheep after treatment with dietary clenbuterol hdrochloride were studied.24 sheep with the weight of(30±5)kg were used to determine the residues and depletion of clenbuterol hdrochloride.Sheep were fed 50 μg/kg clenbuterol hdrochloride for 35 consecutive days.Samples of ovine tissues were collected to determine clenbuterol hdrochloride concentrations by LC-MS/MS.The results showed that residues of clenbuterol hdrochloride were high in ovine eyes.The elimination of clenbuterol hdrochloride was slow in eyes.Clenbuterol hdrochloride concentration in eyes was 42.42 to 63.48 μg/kg on day 14 after withdrawal.The elimination of clenbuterol hdrochloride was the fast in spleen.Clenbuterol hdrochloride was not able to be detected(0.7 μg/kg) in spleen on day 3 after withdrawal.So eyes could be used to monitor and confirm the illegal use of clenbuterol hdrochloride in mutton sheep.