In the present paper, monodisperse poly(lactic acid) (PLA) microspheres (MS) containing the enrofloxacin (ENRO), were manufactured by using a modified solid in oil in water (S/O/W) emulsion solvent evaporation method. In order to prepare PLA microspheres with a higher drug loading efficiency by this modified technique, the test of stability and productivity of the primary emulsion was preliminary examined by change species or concentration of the oil-soluble surfactant and the ratio of water and organic solvent. Firstly, enrofloxacin polylactic acid microspheres (ENRO-PLA-MS) were producted, then the morphology and particle size distribution were estimated by scanning electron microscopy (SEM), its encapsulation efficiency and drug loading efficiency were assessed by High performance liquid chromatography (HPLC). In vivo conditions were simulated by an stable release buffer to obtain a detailed release and polymer degradation profile. Consequently, the ENRO-PLA-MS had a denser structure with a smooth, pore-free surface, the preparation of microspheres was simple, the prepared microspheres had excellent controlled drug release characteristics in vitro.