Background: Callicarpa nudiflora has been usually employed as a traditional Chinese medicine for acesodyne, dispersing edema, and hemostasis; however, its analgesic effects have been poorly considered. Objectives: This article focused on the differences in the contents of seven active chemical constituents in C. nudiflora grown in different regions. Moreover, we examined the analgesic effect of C. nudiflora from five arbitrarily designated regions and studied the relationship between the compound contents and analgesic effect. Materials and Methods: A simple and delicate qualitative tandem liquid chromatography quadrupole time of flight mass spectrometry method was first established for the immediate determination of seven active components (luteolin, apigenin, quercetin, oleanolic acid, ursolic acid, protocatechuic acid, and caffeic acid) in C. nudiflora. Ethanol extracts were prepared from the mature leaves of C. nudiflora from five regions and the writhing test was performed by intragastric administration in mice. SPSS 13.0 and Graph Pad Prism 8.01 software were used to analyze the correlation of all data. Results: Calibration curves presented satisfactory linearity, with correlation coefficients >0.99 for all compounds within the concentration range. The compound contents were uppermost in plants from Hainan Province. The contents of the seven active chemical components inclined in the order of caffeic acid > luteolin > apigenin > oleanolic acid > ursolic acid > protocatechuic acid > quercetin. Pharmacological studies showed that C. nudiflora from all five regions had obvious analgesic effects. Conclusion: These results might be helpful for the screening and cultivation of C. nudiflora and its realistic clinical application.