The invention provides an enrofloxacin injection, comprising following components: 2.5wt%-10wt% of enrofloxacin, 2wt%-10wt% of nicotinamide, 10wt%-30wt% of an organic solvent, 0.01wt%-0.05wt% of disodium ethylenediamine tetraacetic acid, 0.1wt%-0.5wt% of an antioxidant, 0.3wt%-1.4wt% of a pH regulator, and the balance water for injection. After being heated, part of the water for injection is mixed with the enrofloxacin, the nicotinamide, the organic solvent, the disodium ethylenediamine tetraacetic acid and the antioxidant, a mixture is obtained after dissolving, the mixture is mixed with the remaining water for injection after pH of the mixture is adjusted with the pH regulator, and the enrofloxacin injection is obtained. By using the enrofloxacin injection prepared, the technical problem that the enrofloxacin injection has poor stability at low temperature is solved, full play of the therapeutic effect of the enrofloxacin injection is facilitated, the enrofloxacin injection can pass the detection on the quality standard of enrofloxacin injections in Chinese veterinary pharmacopoeia 2010 and is a GB product, formulation cost is low, a production process is easy to operate, and the enrofloxacin injection is suitable for large-scale production in enterprises.