As a semisynthetic tetracycline derivative, doxycycline (α-6-deoxy-5-hydroxytetracycline) is a time-dependent bacteriostatic agent. It is being widely used in the broiler husbandry in China. In this study, doxycycline was orally administered by gavage to 10 healthy broiler chickens at the dose of 20 mg/kg body weight every 24 hr for five total treatments. Plasma samples were collected from each animal at 5, 10, 20, and 30 min and 1, 2, 4, 6, 8, and 12 hr after the first dose, at 0.25, 0.5, 0.45, 1, 2, 3, 4, 6, 7, 13.5, 24, 36, 48, 60, and 72 hr after the last dose. Additional plasma samples were collected at a 24-hr interval during the dosing period (immediately prior to each oral administration). The doxycycline concentrations were determined by high-performance liquid chromatography with an ultraviolet detector and subjected to noncompartmental analysis. Then, the pharmacokinetics profiles were compared after the first and last oral doses. After the first dosing, the elimination half-life, area under the concentration-time curve from 0 hr to ∞, peak concentration, time to reach peak concentration, and volume of distribution per fraction absorbed were determined as 7.78 hr, 94.19 μg·hr/ml, 5.65 μg/ml, 3.50 hr, and 2,502.65 ml/kg, respectively, while the corresponding values of these parameters after the last dose were 19.90 hr, 121.08 μg·hr/ml, 5.71 μg/ml, 7.25 hr, and 5,285.28 ml/kg, respectively. After multiple oral doses, the absorption and elimination both became slower, while the distribution was more extensive than that following a single dose. However, after multiple oral doses, accumulation of doxycycline in plasma was not observed with an average accumulation factor of 1.11.