Integrated chemical and biological characterization of Hypericum perforatum extract using LC-MS/MS and in vitro functional assays

Sci Rep. 2026 Jan 27;16(1):6276. doi: 10.1038/s41598-026-36793-8.

Abstract

Hypericum perforatum L. (St. John's wort) is a medicinal plant known for its diverse secondary metabolites and pharmacological potential. This study provides a comprehensive evaluation of the phytochemical composition and bioactivities of methanolic extracts from wild H. perforatum. LC-MS/MS analysis identified 36 compounds, including phenolic acids (chlorogenic acid, protocatechuic acid) and flavonoids (rutin, isoquercitrin, quercetin), with the first-time detection of genkwanin, vicenin-2, schaftoside, and afzelin, to the best of our knowledge. The extract demonstrated significant antioxidant activity, with a total phenolic content of 203.04 mg GAE/g DW and strong radical scavenging activity in DPPH (544.78 mg TE/g DW) and ABTS (312.15 mg TE/g DW) assays. In vitro assays showed marked cytotoxicity across multiple cancer cell lines, with IC50 values as low as 3.57 µg/mL in A549 cells, indicating stronger activity compared to cisplatin. Flow cytometric analysis revealed substantial apoptosis induction (up to 53.67% in A549 cells) and G0/G1 cell cycle arrest (76.37% in A549). The extract also exhibited moderate antibacterial activity against E. coli and S. aureus, but no activity against P. aeruginosa or Candida species. These results position H. perforatum as a rich source of bioactive compounds with antioxidant, anticancer, and antimicrobial potential, and suggest further research into its therapeutic applications.

Keywords: Antioxidant activity; Apoptosis; Cell cycle arrest; Cytotoxicity; Hypericum perforatum; LC-MS/MS; Phytochemical composition.

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / pharmacology
  • Antioxidants / chemistry
  • Antioxidants / pharmacology
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Chromatography, Liquid
  • Flavonoids / chemistry
  • Flavonoids / pharmacology
  • Humans
  • Hypericum* / chemistry
  • Liquid Chromatography-Mass Spectrometry
  • Microbial Sensitivity Tests
  • Plant Extracts* / chemistry
  • Plant Extracts* / pharmacology
  • Tandem Mass Spectrometry

Substances

  • Plant Extracts
  • Antioxidants
  • Anti-Bacterial Agents
  • Flavonoids

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