Integrated LC-HRMS and HPLC Profiling of Fourteen Anatolian Hypericum Extracts Reveals Distinct Chemometric and Bioactivity Patterns.

Background/Objectives: Anatolia hosts a rich diversity of Hypericum taxa; however, the chemical and biological properties of most species remain insufficiently characterized. Methods: This study combined liquid chromatography-high-resolution mass spectrometry (LC-HRMS) with high-performance liquid chromatography coupled to diode-array detection (HPLC-DAD) to profile 14 extracts obtained from 12 Anatolian species together with H. perforatum, and to examine whether metabolic variation aligns with bioactivity trends. Results: Chemometric analyses (Principal Component Analysis-PCA-and Hierarchical Cluster Analysis-HCA) revealed distinct chemotypes primarily driven by phenolic acids and flavonol glycosides, with hyperoside emerging as a consistently abundant marker. Phenolic-rich extracts displayed enhanced functional properties in multiple assays. Among them, H. triquetrifolium showed the strongest acetylcholinesterase inhibition and notable antioxidant capacity, whereas the selected taxa exhibited comparatively high tyrosinase or α-glucosidase inhibition. Antimicrobial testing indicated narrow-spectrum activity limited to Gram-positive bacteria and Candida spp. Conclusions: The integration of metabolite profiling with chemometric and functional data highlights clear chemotype-bioactivity associations across Anatolian Hypericum taxa and underscores the analytical value of LC-HRMS-guided chemotyping for pharmacognostic assessments. These results broaden the phytochemical and bioactivity landscape of this genus and provide a comparative framework for prioritizing species for future targeted investigations.