The invention provides a preparation method of phosphamycin levophosphorylamine. The preparation method comprises the following steps: dissolving cis-propenylphosphonic acid in an alcohol solvent at room temperature, slowly dropwise adding (+)-alpha-phenylethylamine, regulating the pH value of the formed system to 5.5-6, continuing stirring for 1-3 minutes, adding a silver catalyst, slowly and dropwise adding hydrogen peroxide, continuing stirring for 10-30 minutes, then rapidly heating the system to 50-55 DEG C, conducting filtering while the system is hot, and then cooling, crystallizing andwashing the filtrate successively to obtain the phosphamycin levophosphorylamine. According to the invention, silver carbonate is used as a catalyst, hydrogen peroxide is used as an oxidizing agent,heating is not needed in an oxidative cyclization process, and a reaction can be performed at normal temperature. The silver carbonate has very high catalytic activity in the invention, and compared with the prior art, the method has the advantages of small dosage, mild reactions, effective shortening of reaction time, simple post-treatment, and realization of separation of the catalyst from the system only through filtration of the system while the system is hot.