Objective The mouse ear blue staining test was used to evaluate anaphylactoid reactions to different injected doses of osteopeptide. Methods The positive model was selected by using histamine phosphate, histamine hydrochloride and histamine as the positive drugs. Mouse ear blue staining tests were performed by injecting different doses of osteopeptide. The rate of blue staining and the rising rate of evans blue were used as the indices for evaluating the anaphylactoid reactions. Results The positive model established by histamine phosphate as the positive drug exhibited obvious anaphylactoid reactions. The osteopeptide injections showed positive reactions to various degrees at high and medium doses and negative reactions at low doses. Conclusions Based on the result of the mouse ear staining test, the anaphylactoid reactions to osteopeptide injections at different doses were intuitively evaluated. These data may be used as a reference for the evaluation of anaphylactoid reactions of other complex matrix preparations.