Electrochemical removal of synthesis solution containing 50 mg/L auramine-O through Ti/PbO2 electrodes and carbon felt air-diffusion electrode to in situ generate hydrogen peroxide (H2O2) by the oxygen (O2) reduction. The effects of operating conditions, FeSO4·7H2O concentrations, current density, initial concentration, initial pH value and sodium sulfate (Na2SO4) concentration were investigated. For reaction of 60 min, the auramine-O removal rate reached 99% and total organic carbon (TOC) removal rate was close to 64.27%. This means that the organic macromolecules in the pollutants have been completely destroyed and broken down. The degradation of auramine-O under electro-Fenton (EF) process using Ti/PbO2-carbon felt was analyzed from the dimensions of TOC, mineralization current efficiency, energy consumption, degradation mechanism and kinetics. Comparing EF-Ti/PbO2 with Ti/PbO2 anode, AO (anodic oxidation)-Ti/PbO2 anodic oxidation using Ti/PbO2 anode and EF-Pt EF process with Pt anode, the consequences demonstrated that Ti/PbO2-EF process by hydrogen peroxide produced in situ was the best choice than others.