Objective To analyze some potential factors which might affect the active systemic anaphylaxis (ASA) test on guinea pig. Methods In GLP conditions, guinea pigs from 300 to 400 grams were used as experimental subjects. By using the materials including bovine serum albumin and saline which are recommended for the control subjects by the guidelines, some experimental design factors such as sensitizing dose, challenge dose, challenge interval and challenge injection route, and the frequency of challenge were studied during the systemic anaphylaxis in guinea pigs. ResultsIn a certain dose range, the incidence of allergic reactions in guinea pigs was not related with systemic sensitizing dose, challenge dose and challenge injection approach, but the severity of anaphylaxis symptoms has an obvious relationship with sensitizing dose, challenge dose and challenge injection route. Conclusion Study on the drug-induced anaphylaxis should refer to the guidelines and pay more attention to experimental design dose and the route of administration.