Comparative Antioxidant Profiling of Medicinal Mushrooms Collected in Serbia: LC‐MS/MS Characterization and In Vivo Oxidative Stress Model

This study explores the in vitro antioxidant properties of hydroethanolic (70% EtOH), hot water (H 2 O), and chloroform (CHCl 3 ) extracts obtained from Cyclocybe aegerita , Ganoderma lucidum , and Pleurotus ostreatus , along with their mycochemical profiles determined by LC‐MS/MS and total phenolic content (TP). In addition, the in vivo antioxidant potential was specifically evaluated using the 70% EtOH extracts in a CCl 4 ‐induced oxidative stress model. LC‐MS/MS analysis identified p ‐hydroxybenzoic, quinic, gentisic, p ‐coumaric, and cinnamic acids, with cinnamic acid being most abundant in C. aegerita (>1500 μg/g d.e.). The highest TP was found in C. aegerita H 2 O and 70% EtOH extracts. In vitro antioxidant assays revealed that G. lucidum H 2 O extract exhibited the strongest DPPH scavenging (IC 50 = 37.52 ± 0.86 μg/mL), P. ostreatus 70% EtOH extract showed the highest ABTS activity (354.10 ± 4.80 mg TE/g d.w.), while C. aegerita demonstrated the most potent hydroxyl radical scavenging. In vivo, P. ostreatus significantly mitigated CCl 4 ‐induced oxidative stress by preserving catalase (CAT), glutathione (GSH), and peroxidase (Px) activities, while reducing lipid peroxidation (LPx) and xanthine oxidase (XOD) levels. These findings highlight the species‐specific antioxidant potential of G. lucidum and C. aegerita , with P. ostreatus demonstrating particular promise for biotechnological applications targeting oxidative stress. Given their rich polyphenolic content, these mushrooms represent potential candidates for the development of functional foods, nutraceuticals, or pharmaceutical formulations.