Abstract: To establish an immunoassay method on salidroside (SAL), it is necessary to firstly create a complete SAL antigen. This study applied the sodium periodate oxidation method to synthesize the immunizing antigen, SAL-BSA, and the coating antigen, SAL-OVA, which were positively identified by means of ultraviolet spectrum and SDS-PAGE. Subsequently, lab mice were immunized with the SAL-BSA to produce antiserum against SAL. The antibody titer was, then, determined by indirect ELISA. The results showed that SAL was conjugated with BSA/OVA in a ratio of 7:1, and the serum antigen titer reached 1:4 000. With the complete SAL antigen successfully synthesized, the preparation of monoclonal antibodies for the establishment of an applicable immunoassay methodology became possible.