Objective  To prepare monoclonal antibody of salidroside for the development of an indirect competitive enzyme linked immunosorbent assay (ic-ELISA) methodology to accurately and rapidly detect salidroside in medicinal materials.

  Method  Balb/c mice of 6-8 weeks old were immunized with the prepared salidroside-BSA. Specificity of the anti-serum was determined by ELISA. Spleen cells from the mice with positive result were fused with SP2/0 myeloma cells. Monoclonal hybridoma cells were screened by indirect ELISA with a limited dilution. Ascitic antibodies were induced and prepared from the positive cell line. Specific antibody against salidroside was used to establish ic-ELISA detection method that was verified by its specificity, precision and recovery rate on a known standard.

  Result  The sensitivity of ic-ELISA was 49.33 ng·mL^-1 with a liner range of 4.07-598.45 ng·mL^-1. The detection limit of the method was 1.77 ng·mL^-1. The newly developed method detected salidroside in spiked samples with a recovery rate of 94.3% within the designed range. The relative standard deviation of the measurements was less than 4.5% on both intra- and inter-day assays. The determination by ic-ELISA agreed with that obtained by HPLC.

  Conclusion  The newly established ic-ELISA method based on the salidroside monoclonal antibody was considered appropriate for the detection.