Influence of Extraction Method on the Phytochemical Composition and Biofunctional Potential of <i>Centaurea urvillei</i> subsp. <i>Stepposa</i> : A Multi‐Assay and In Silico Perspective
Zeyneb Karakuş
Abstract
ABSTRACT This study explores how extraction methods influence the phytochemical composition and bioactivity of Centaurea urvillei subsp. stepposa , an Anatolian endemic. Methanolic extracts were obtained via maceration (MAC‐ME), Soxhlet (SOE‐ME), and ultrasound‐assisted extraction (UAE‐ME). LC‐MS/MS profiling identified chlorogenic acid as the dominant phenolic (up to 2181 µg/g in MAC‐ME), while UAE‐ME enriched glycosylated flavones such as apigenin 7‐glucoside (862 µg/g). Although total phenolic and flavonoid contents were similar across extracts, antioxidant and enzyme inhibition assays revealed method‐specific differences. UAE‐ME showed the highest Relative Antioxidant Capacity Index (RACI = +0.31), with strong metal‐chelating and phosphomolybdenum activities. MAC‐ME exhibited superior AChE inhibition (IC 50 = 1.24 mg/mL), while UAE‐ME best inhibited α‐amylase (IC 50 = 1.74 mg/mL). Molecular docking and MM/GBSA analyses confirmed strong interactions between chlorogenic acid, apigenin 7‐glucoside, and enzyme targets (Δ G _MM/GBSA as low as –48.54 kcal/mol). Correlation analyses highlighted flavone‐type compounds as major contributors to bioactivity. The results underscore the impact of extraction technique on biological potential and identify UAE as suitable for generating a relatively balanced bioactive profile. This work highlights C. urvillei subsp. stepposa as a botanically rich source of phenolic compounds with potential relevance for future nutraceutical and cosmeceutical research.