To reveal the characteristics of tetracycline (TC) photocatalytic degradation under Cu(II) coexistence, effects of Cu(II) on TC photocatalytic degradation by ZnO nanoparticles (ZnO NPs) as a function of pH, humic acid (HA), and initial Cu(II) concentration were investigated. Interaction of TC with Cu(II) in the treatment process was analyzed by circular dichroism (CD) spectroscopy, while TC degradation pathway was investigated by high-performance liquid chromatography-mass spectrometry. Sixty-five percent and ninety-one percent TC degradation within 60 min in the absence and presence of Cu(II), respectively, was reported. Both adsorption and photocatalytic degradation of TC under Cu(II) coexistence increased with increasing pH from 3 to 6, while decreased with further increase in pH. HA inhibited the degradation of TC by ZnO NPs both in the presence as well absence of Cu(II), while TC degradation decreased from 91 to 73% and from 73 to 37% in the presence and absence of Cu(II), respectively. TC degradation by ZnO NPs first increased then decreased with increasing Cu(II). Maximum TC degradation (about 94%) was obtained in the optimum concentration range of Cu(II) (0.05-0.15 mmol/L). In addition, there was a lag effect between TC adsorption and degradation on ZnO NPs. TC degradation was improved via Cu(II)-TC surface complexation and followed N-demethylation and hydroxylation routes. This study could be of potential importance in extrapolating the transformation of TC or other antibiotics under the coexistence of heavy metals in water.