Abstract: Antibacterial materials were extracted from Medicago falcata L. and the inhibitory effect of the extract on different test strains was evaluated. Single-factor and orthogonal test experiments were used to explore the optimal extraction processing parameters for antibacterial materials from Medicago falcata L. using Bacillus subtilis, Escherichia coli, Staphylococcus aureus and Proteus vulgaris as the indicators. The optimal extraction processing parameters for antibacterial materials from Medicago falcata L. were ethanol concentration of 80%, material-liquid ratio of 1∶12 and extraction time of 2.0h. The extract obtained by optimal extraction enhanced inhibitory effect on test bacterial strains.