Abstract: AIM: To observe the effects of C48/80 on RBL-2H3 cell degranulation orthogonal test, optimize the experimental conditions and discuss the possible detection methods of anaphylactoid reaction . METHODS: The β-hexosaminidase assay of RBL-2H3 cells incubated with C48/80 were tested by orthogonal test considering two laboratorial factors time and cell concentration.According the results the optimized cell model in vitro were set up and disscus the superior and the inferior aspects between different detection means.RESULTS: Different concentrations and time could make significance difference on RBL-2H3 cell model with degranulation.CONCLUSION: Anaphylactoid reaction based on RBL -2H3 cell model in vitro could be built up on the cell concentration of 2×10⁵ under 60 minutes incubation time , which may be appropriate conditions in order to get stable cell model because the degranualtion was significantly increased and dose-dependent under this condition.

Key words: Anaphylactoid reaction, RBL-2H3 cell, Degranulation, Orthogonal test