This study quantifies the rapid removal of tetracycline (TC) in acetic acid solution by ozonation after extraction from water into acetic acid solution using granular activated carbon (GAC). Systematic laboratory experiments show that GAC can be used as an adsorbent to transfer TC from water to acetic acid. Ozone gas has a high degree of stability and solubility in acetic acid, and therefore apparently enhances the removal rate of TC. TC removal efficiency is better in 10% acetic acid solution than that in water, 15% and 20% acetic acid solution. Removal rate of TC in the acetic acid solution decreases as the initial molar ratio of TC and O-3 increases. Inhibition effect of tertiary butyl alcohol (TBA) on TC removal is smaller in acetic acid than that in water. metal-TC complex was produced in the presence of Fe3+ and Co3+, which inhibited the degradation of TC by ozone in acetic acid. Removal of TC by ozone in the acetic acid solution might be affected by multiple factors, which should be considered in practical applications.