Sinusoidal alternating electro-Fenton (SAEF) is a new type of advanced electrochemical oxidation technology for the treatment of refractory organic wastewater. In this research, the removal performance and degradation mechanism of tetracycline (TC) were investigated, and the optimal operation parameters were determined. Scanning electron microscope (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectrometer (FTIR) were used to characterize the morphology, elemental composition, crystal structure, function groups of sludge produced by SAEF. UV-visible spectroscopy (UV) and liquid chromatograph-mass spectrometer (LC-MS/MS) were employed to determine the concentration of organic matter, middle products of decomposed organics in the SAEF process, respectively. The results showed that the removal rates of TC, chemical oxygen demand (COD), electric energy consumption (EEC) and the amount of produced sludge (Ws) are 94.87%, 82.42%, 1.383 kWh⋅m−3 and 0.1833 kg⋅m−3 by SAEF, respectively, under the optimal conditions (pH = 3.0, conductivity (κ) = 1075 μS⋅cm−1, current density (j) = 0.694 mA⋅cm−2, initial c (TC) = 100 mg·dm−3, c [30%H2O2] = 1.17 cm3⋅dm−3, frequency (f) = 50 Hz, t = 120 min). Compared with pure direct electro-Fenton (DEF) or sinusoidal alternating current coagulation (SACC), SAEF was a highly effective method with low-cost for the treatment of TC wastewater. It was found that the conjugated structure of TC was destroyed to generate intermediate products, and then most of them was gradually mineralized into inorganic materials in the SAEF process.