Backgrounds and Objectives: Antibiotics can’t be removedefficiently by conventional wastewater treatment. Nano Zero Valent Iron (nZVI) has high efficiency of degradation of contaminants such as heavy metal, nitro aromatic compounds and insecticides. Ozonation is one of the most well known methods for remove of chemichal-resistant materials like antibiotics of water and wastewaterMaterials and Methods: This study is experimental. nZVI and ozonation were used to remove of penicillin G. for determination removal efficiency of penicillin, COD and TOC, factors: initial pH, dose of nano particle, initial penicillin dose, reaction time with nano particle and ozone were investigated.Results: Removal efficiency of penicillin G, COD, TOC in stage of reaction with nZVI were 96.35%, 73.13% and 47.45%, respectivly with reaction time: 60 min, initial penicillin concentration: 100 mg/L, COD: 1429 mg/L, TOC: 510 mg/L, pH=7, nano particle iron concentration: 0.25 g/L. In the stage of reaction with ozone, removal efficiency of penicillin G, COD and TOC were 99.5%, 96.35% and 92.65%, respectivly with reaction time: 13 min, pH=9, initial penicillin 20 mg/l, COD: 356 mg/L, TOC: 136 mg/L, volumetric flow rate of ozone: 5 L/min.Conclusion: The best removal efficiency in stage of nano particles reaction was in pH=7, initial nZVI concentration 0.25 g/L, initial penicillin concentration 100 mg/L and time about 60 min. The best removal efficiency in stage of ozonation was in pH=7 and time about 9 min.