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However, the active ingredients are unclear. This study aimed to identify and validate the active ingredients of MC. UPLC fingerprints of 23 batches of MC from various origins were analyzed. The activating blood efficacy of MC was assessed by evaluating the inhibitory effects on thrombin and factor Xa (FXa) using the chromogenic substrate method. Active ingredients were identified through spectrum\u2010effect relationship analysis using gray relation analysis (GRA), partial least squares (PLS), and support vector machine (SVM) algorithms. Consequently, five components were identified as potential active ingredients: mudanpioside H, oxypaeoniflorin, 1,2,3,4,6\u2010penta\u2010O\u2010galloyl\u2010\u03b2\u2010d\u2010glucose (PGG), benzoyloxypaeoniflorin, and suffruticoside A\/B\/C\/D. The pharmacological activities of these five active ingredients were further confirmed by measuring their thrombin inhibition ability and antithrombotic effects in zebrafish, and their interactions with thrombin and FXa were examined using molecular docking technology. Oxypaeoniflorin, benzoyloxypaeoniflorin, and PGG demonstrated significant efficacy in promoting blood circulation and resolving blood stasis, as well as strong binding affinities. This study provides a biochemical foundation for the anticoagulant effects of MC and offers valuable insights for quality control and the development of novel anticoagulant ingredients drugs.<\/jats:p>","DOI":"10.1002\/bmc.70060","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T04:21:26Z","timestamp":1742185286000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Identifying of Anticoagulant Ingredients From Moutan Cortex Based on Spectrum\u2010Effect Relationship Analysis Combined With GRA, PLS, and SVM Algorithms"],"prefix":"10.1002","volume":"39","author":[{"given":"Weijie","family":"Pan","sequence":"first","affiliation":[{"name":"School of Traditional Chinese Medicine Guangdong Pharmaceutical University\/Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica, State Administration of Traditional Chinese Medicine (TCM)\/Engineering Technology Research Center 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