Bioactivity assessment and LC–MS/MS-based phenolic characterization of Gypsophila pilosa

In this study, the antioxidant, anticholinesterase, antiproliferative activities, and phenolic compound profile of ethanol extracts of Gypsophila pilosa were comprehensively evaluated. Plant samples were collected from Şanlıurfa province, Türkiye, and after shade-drying under laboratory conditions, they were extracted with ethanol using a Soxhlet apparatus at 50 °C. The antioxidant capacity results revealed that total antioxidant status was 3.528 ± 0.085 mmol/L, total oxidant status (TOS) was 10.248 ± 0.072 µmol/L, and oxidative stress index was 0.291 ± 0.009. In addition, DPPH radical scavenging activity and ferric reducing antioxidant power (FRAP) were determined as 92.317 ± 2.336 mg TE/g and 128.016 ± 2.480 mg TE/g, respectively, indicating a strong antioxidant potential. In anticholinesterase assays, the extract exhibited inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with IC₅₀ values of 160.163 ± 2.155 µg/mL and 223.903 ± 1.049 µg/mL, respectively. The antiproliferative activity was evaluated on A549 and MCF-7 cancer cell lines, as well as the HaCaT normal cell line. The extract demonstrated a dose-dependent reduction in cell viability in cancer cell lines, while comparatively lower cytotoxic effects were observed in normal cells at lower concentrations, suggesting a degree of selective activity. LC-MS/MS analysis revealed the presence of several bioactive phenolic compounds, including catechin hydrate, caffeic acid, gallic acid, and kaempferol, which are known for their antioxidant and cytotoxic properties. These compounds may contribute to the observed biological activities through overlapping and complementary mechanisms. In conclusion, G. pilosa exhibits notable antioxidant, enzyme inhibitory, and antiproliferative activities, which may be associated with its phenolic composition. However, further studies are required to clarify its selectivity and underlying mechanisms before potential pharmaceutical applications can be considered.