Integrated Phytochemical Profiling and Multifunctional Bioactivities of <i>Bellis annua</i> Extracts Obtained by Distinct Extraction Techniques

This study presents an integrated chemical and biological assessment of Bellis annua methanolic extracts obtained through maceration (MAC), Soxhlet extraction (SOE), and ultrasound-assisted extraction (UAE). LC-electrospray ionization-mass spectrometry (ESI-MS)/MS profiling enabled the quantification of major phenolic compounds, including chlorogenic acid, hyperoside, hesperidin, and several hydroxycinnamic acids, revealing extraction-dependent variations in phenolic distribution. While total phenolic levels were comparable across methods, UAE yielded the highest flavonoid content and a broader enrichment of glycosylated flavonoids and hydroxycinnamates. Antioxidant capacity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging assays, ferric reducing antioxidant power, cupric reducing antioxidant capacity, the phosphomolybdenum method, and a metal-chelation assay. All assays indicated notable activity of the extracts. UAE exhibited the strongest overall antioxidant performance as indicated by the Relative Antioxidant Capacity Index. Enzyme inhibition assays revealed method-specific differences: MAC showed the most pronounced acetylcholinesterase and α-amylase inhibition, whereas SOE demonstrated superior tyrosinase inhibition. Correlation analyses further indicated that flavonoid-rich extracts were strongly associated with radical-scavenging and metal-chelating activities, while specific phenolic acids contributed differentially to enzyme inhibition patterns. These findings highlight the influence of extraction technique on the chemical composition and multifunctional bioactivity of B. annua, supporting its potential as a promising natural source of phenolic-driven antioxidant and enzyme-inhibitory agents with relevance to pharmaceutical and nutraceutical applications.