Nano micro-electrolysis materials (nMETs) have been used to degrade refractory pollutants in batch experiments. The reasonable formation mechanism of nMETs was given through DMXY digital biomicroscopy. Based on the kinetic data of Chlortetracycline (CTC) removal by nMETs in batch experiments, combined with the binomial distribution equation of CTC reduction by nano materials an experimental-scale fluidized bed (ESFB) was designed. The effects of CTC removal performance, pH and iron ion concentration were investigated. Under pure CTC solution environment, the experimental data showed that the average removal rates of CTC by nMET and nano micro-electrolysis material with loading copper (Cu-nMET) are 90.0% and 95.7% in ESFB, respectively. In the presence of nitrate, although the consumption of two kinds of nano-materials increased, their removal efficiencies of pollutants have 2.2%, 0.2% increase compared with the nitrate-free ESFB. At the same time, the CTC degradation pathway and the enhanced removal mechanism by Cu-nMET was proposed. Through microelectrolysis reaction, complexation reaction and the active substances produced, the intermediate products can be degraded completely to NH, CO, HO and so on. This study aims to provide a theoretical basis for the environmental application of nMETs.