INTRODUCTION: Myricariae Ramulus (MR) is a traditional anti-inflammatory Tibetan medicine derived from the branches and leafy twigs of various Myricaria plants, such as Myricaria wardii Marquand. OBJECTIVE: This study performed spectrum-effect analyses on 15 batches of MR, sourced from various origins and medicinal parts, to identify quality markers associated with its anti-inflammatory effects. MATERIALS AND METHODS: The anti-inflammatory effects of different extracts and fractions from M. wardii were evaluated in lipopolysaccharide-infected mice. Further analysis was conducted utilizing UPLC-Q-TOF-MS/MS in the negative ion mode to recognize the compound composition of ethyl acetate (EA) fractions from the branches and leafy twigs of M. wardii. Subsequently, anti-complementary and antioxidant activities were conducted via the hemolysis and ABTS radical scavenging methods, and UPLC chromatograms for 15 batches of samples were established. Finally, spectrum-effect analysis was performed by gray relational analysis and partial least squares regression to identify the quality markers. RESULTS: The EA fraction effectively alleviated inflammatory responses by regulating IL-6 and myeloperoxidase. A total of 46 compounds from M. wardii were recognized, which mainly encompass gallic derivatives, phenolic amides, flavonoids, and terpenoids. Activity assays demonstrated that the two activities of leafy twigs were significantly stronger than those of branches. Moreover, an obvious distinction in chemical composition between the two parts was observed. Combining the spectrum-effect analysis results, methyl 3,4-dihydroxy-5-methoxybenzoate, myricarin A, protocatechualdehyde, and N-feruloyl normetanephrine from the branches, and protocatechuic acid from the leafy twigs were considered quality markers. CONCLUSION: These results provide insights into improving the quality control of MR.