Activated carbon enriched with high concentrations of gentamicin (ACG) was generated in the production process of gentamicin. Inappropriate handling methods for ACG not only squanders carbon resource, but also seriously hinders achieving global carbon neutrality and hazardous to human health. In the present work, thermal and carbon co-activated persulfate method (TC-PS) was developed to regenerate ACG with degrading gentamicin. The results showed that ACG was effectively regenerated by TC-PS, restoring the adsorption performance for gentamicin. When the treatment temperature was 80 °C, the persulfate dosage was 20 mM and the initial pH was 3.0, the degradation efficiency of gentamicin reached 100%. The HO• and SO4•- were the major reactive species for gentamicin degradation. The possible degradation routes of gentamicin were proposed and the safety assessment indicated that the produced intermediates during the regeneration process of ACG by TC-PS have insignificant impact on the biological and ecological environment.