Abstract: OBJECTIVE To prepare mogrosides V (MogV) specific antiserum and establish the competitive-inhibitive ELISA method to detect MogV content. METHODS Antiserum was prepared with the V-Mogrosides succinate-bovine serum albumin(MogV-HS-BSA) as the immunogen (ratio about 44∶1), the competitive inhibition ELISA method for MogV content was established with the V-Mogrosides succinate-human serum protein (MogV-HS-HSA) with a ratio of about 64∶1 as coating antigens. RESULTS The coating antigen working concentration was 1 μg·mL^-1, the best serum dilution is 1∶8 000. Within MogV concentration 1×10^-4-1 μg·mL^-1, the regression equation was Y=-0.158X+0.298, r=0.991 0. CONCLUSION MogV anti-serum is successfully prepared. Within 1×10^-4-1 μg·mL^-1, the MogV concentration and OD value(405 nm) of the antiserum shows a good linear relationship.