Abstract: Objective To analyze the impurity spectra of betahistine hydrochloride injection to provide a basis for quality control of this variety. Methods HPLC-QE-MS technology were applied to identify and analyze the structure of an unknown impurity of betahistine hydrochloride injection. According to forced degradation test, the source of impurities and impurity spectra were determined. ADMET Predictor software was used to predict the toxicity of impurities. Results The method established for related substances was simple with high accuracy and good reproducibility, total of four unknown impurities were detected, Impurities 1, 2, and 3 were all process impurities, impurities A, impurities B, and impurities 4 were all degradation impurities, and impurity C was both process and degradation impurity. The toxicity prediction results showed that impurity 1, 2, and 3 had potential acute toxicity in rats; Impurities A, B, 1, and 4 had potential liver toxicity. Correlation analysis of impurity mass spectrometry and sterilization process showed that betahistine hydrochloride was thermally unstable, and the sterilization process currently used by various enterprises would increase the quantity and content of related substances. Conclusion By studying the impurity mass spectrometry of betahistine hydrochloride inject

Key words: betahistine hydrochloride injection, impurity spectrum, genetic toxicity, HPLC-QE-MS, quality control, methodology