Utilizing ultra-performance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry (UPLC-Q/Orbitrap HRMS), in conjunction with network pharmacology and zebrafish experimental validation, this study aimed to elucidate the potential mechanisms and active components of Brassica rapa L. in the treatment of hyperuricemia (HUA). Initially, various extracted fractions of B. rapa were screened for their xanthine oxidase inhibitory activity in vitro. The fraction exhibiting the strongest inhibitory effect was identified, and its chemical constituents were analyzed by UPLC-Q/Orbitrap HRMS. Leveraging network pharmacology, we further explored the potential therapeutic mechanisms for HUA. Subsequently, zebrafish experiments were conducted to validate the selected targets. The findings indicated that the most effective uric acid-lowering fraction was the 70% ethanol extract of B. rapa, containing 76 identified compounds, 37 of which were potentially bioactive. These active ingredients primarily functioned through the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway and others. Eight key targets, including adenosine triphosphate-binding cassette transporter protein G2 (ABCG2),organic anion transporter 1 (OAT1), and urate anion transporter 1 (URAT1), along with nine core components like sinensetin, palmitic acid, and phenylethyl 2-glucoside, were identified. Molecular docking revealed a high affinity between the core components and the key targets.Final validation through zebrafish experiments demonstrated that the 70% ethanol extract significantly upregulated the expression of OAT1, a core target exhibiting the highest degree value among HUA-associated targets. Overall, this study established a foundation for exploring the uric acid-lowering properties of B. rapa, uncovered its preliminary mechanisms in treating HUA, and provided a theoretical framework for future investigations.