The invention discloses a method for identifying deer hard antler button effective components for treating mastitis. The method comprises the following steps: S1, setting deer hard antler button extracts with gradient concentration; S2, determining and screening the change level of inflammatory cytokines; S3, determining and screening out an effect peak value, and collecting a treating fluid corresponding to the effect peak value; S4, verifying a single-component peak, and determining the treating fluid with a verified effect peak value; and S5, determining chemical components of the effective components. According to the invention, deer hard antler button is selected, and detailed components are obtained through pretreatment, concentration gradient setting, immunoassay, enzyme-linked immunosorbent assay, real-time fluorescent quantitative PCR detection, immunoblotting analysis, fingerprint map, grey correlation analysis, verification of a single-component peak and non-targeted metabonomics of a liquid chromatograph-mass spectrometer, so that comprehensive judgment of efficacy and accurate identification are realized.