This study aimed to establish high performance liquid chromatography(HPLC) fingerprints of Rhinacanthus nasutus(L.) Kurz from different origins and use multimodal chemical stoichiometry analysis to evaluate their quality.Methods:Ten batches of HPLC fingerprints of Rhinacanthus nasutus(L.) Kurz were established，and the quality of Rhinacanthus nasutus(L.) Kurz from different origins was evaluated by using similarity evaluation，clustering analysis(CA)，and orthogonal partial least squares-discriminant analysis(OPLS-DA).Results:The fingerprints identified 10 common peaks through separation and purification，including 12-hydroxylapachenole，rhinacanthin-A，rhinacanthin-E，heliobuhthalmin，2,2-Dipheny-2H-benzo chrome，rhinacanthin-D，rhinacanthin-Q，rhinacanthin-B，rhinacanthin-C，and rhinacanthin-N.The similarity evaluation results showed that except for the samples from Nanning，Yongning，Pubei，and Pingxiang in Guangxi，the similarity among the 10 batches of samples was greater than 0.900 0.By combining multimodal chemical stoichiometry analysis，the 10 batches of samples were divided into two categories，and characteristic biomarkers that may lead to differences in the quality of Rhinacanthus nasutus(L.) Kurz were identified as rhinacanthin-C and rhinacanthin-A.Conclusion:The fingerprint used in this study is simple and has good repeatability.When combined with multimodal chemical stoichiometry analysis，it can effectively evaluate the quality of Rhinacanthus nasutus(L.) Kurz medicinal materials from different origins.