Molecular Networking-Guided Phytochemical Profiling and Anti-Inflammatory Evaluation of Honglanqi, an Underutilized Commercial Specification of Astragali Radix

Honglanqi (HLQ), a distinctive commercial specification of Astragali Radix (AR) characterized by rhytidome and interxylary cork, remains undervalued despite originating from premium long-cultivated material. The ethyl acetate fraction (HLQE) showed the greatest anti-inflammatory activity among the polarity-based fractions of an HLQ ethanol extract. We applied an integrated ultra-high performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC–QTOF-MS/MS) and feature-based molecular networking (FBMN) strategy to systematically profile the chemical composition of HLQE. A total of 102 metabolites were annotated across seven compound classes, of which 18 were predicted as potential new compounds, and 37 were reported in AR for the first time. Three undescribed compounds, astragalinin A, astragalinin B, and astragquinone, were isolated and characterized, together with 19 known compounds. In LPS-stimulated RAW264.7 macrophages, astragalinin A exhibited potent inhibition of TNF-α and IL-6 (IC50 < 10 μM). Astragquinone showed significant TNF-α inhibition with a favourable safety profile. Stereochemical configuration critically influenced anti-inflammatory potency, as demonstrated by the enantiomeric pair. Molecular docking predicted favourable binding orientations toward TNF-α and IL-6 in silico. These findings provide a comprehensive phytochemical and bioactivity profile of HLQ, supporting its value-added utilization as an underutilized AR resource.