Direct spray mass spectrometry was used to simply and rapidly differentiate Mutong of Aristolochiaceae from other two kinds of Mutong medicinal materials (Lardizabalaceae and Ranunculacea) by analyzing the chemical profile of Mutong of Aristolochiaceae.A novel method for determination of magnoflorine content in Mutong of Aristolochiaceae was established.The results showed that Mutong of Aristolochiaceae could be identified according to the symbolic component, magnoflorine.Under positive ion mode, semi-quantitative result based on the signal strength ratio of magnoflorine and nuciferin was obtained by choosing nuciferin as an internal standard.The method showed good linear coefficient in the concentration range of 0.50-20.00 mg/L of magnoflorine.The limit of detection was 0.1 mg/L.The method was simple and fast, and could be used for direct and rapid in-situ analysis and identification of Mutong of Aristolochiaceae from other closely related Mutong herbal species without sample pre-treatment.The results were important for the quality control of Mutong herbal medicine.