An integrated NP-CMC-HPLC strategy for screening bioactive candidates with antidepressant potential and quality characterization of Curcuma wenyujin.

Depression is a complex disorder involving multiple pathological processes, and the identification of bioactive candidates from traditional Chinese medicine (TCM) remains challenging. This study established an integrated network pharmacology (NP)-cell membrane chromatography (CMC)-high-performance liquid chromatography (HPLC) strategy (NP-CMC-HPLC) for activity-guided screening of potential bioactive compounds and activity-associated quality characterization of the dried tuberous root of Curcuma wenyujin Y. H. Chen & C. Ling (WYJ). The chemical profile of WYJ was comprehensively characterized using ultra-performance liquid chromatography coupled with quadrupole Orbitrap mass spectrometry (UPLC-Q-Orbitrap-MS), combined with headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) and headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS). A total of 184 compounds were identified, and potential bioactive compounds were prioritized through network pharmacology analysis. Multi-target cell membrane chromatography (CMC) systems based on 5-hydroxytryptamine 1 A receptor (5-HT1A), 5-hydroxytryptamine 2 A receptor (5-HT2A), dopamine D1 receptor (DRD1), dopamine D2 receptor (DRD2), and monoamine oxidase A (MAO-A) were subsequently established, enabling the identification of six compounds exhibiting target-associated retention behavior. Molecular docking and a corticosterone-induced PC12 cell injury model provided complementary computational and cellular evidence supporting compound selection. Finally, a parallel reaction monitoring (PRM)-based method was developed for the quantitative analysis of four non-volatile CMC-prioritized compounds. The proposed strategy integrates chemical profiling, target affinity screening, cellular evaluation, and quantitative analysis to facilitate the discovery of bioactive candidates and activity-associated quality characterization of complex herbal medicines.