Higenamine (HM), classified as an S3 β2 agonist and prohibited by the World Anti-Doping Agency (WADA), is prevalent in spices and traditional Chinese herbal medicines.Accidental consumption can lead to HM positivity in athletes, necessitating daily monitoring. However, current detection strategies predominantly involve instrumental analysis, with limited development of rapid methodologies incorporating sample preparation for practical, daily applications. In this study, a novel method has been developed for the rapid and quantitative detection of HM in throat lozenges, plant beverages, syrups and grass coral. This innovative approach combines the use of immunomagnetic beads and an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to achieve efficient enrichment and detection. In the assay development, a highly sensitive anti-HM monoclonal antibody 1E9 (1E9-mAb) was generated and its binding mechanism to HM was investigated by theoretical simulations. Under optimum conditions, the sensitivity (IC