LC‐HESI‐MS/MS Profiling and Evaluation of Antioxidant, Anti‐Inflammatory, Anti‐α‐Amylase, and Anti‐Acetylcholinesterase Activities of Rhus tripartitum L. Leaf Extract: In Vitro and In Silico Studies

ABSTRACT This study aims to assess the biological characteristics of Rhus tripartitum leaves and identify their promising biomolecules. The numerous extracts of R. tripartitum leaves were evaluated, in vitro, for their phytochemical profile and antioxidant, anti‐acetylcholinesterase, α‐amylase, and anti‐inflammatory activities. The methanolic and ethyl acetate extracts showed the highest levels of phenolics, flavonoids, and tannins, and exhibited the strongest antioxidant effects in DPPH, FRAP, and TAC assays. Regarding the ethyl acetate extract, it demonstrated the greatest in vitro enzyme inhibitory activity against acetylcholinesterase (IC 50 = 0.186 mg/mL), α‐amylase (0.100 mg/mL), and anti‐inflammatory (0.151 mg/mL), respectively. Using LC‐HESI‐MS/MS analysis, 28 compounds were characterized, including phenolic acids, flavonoids, and fatty acids, notably quercetin and kaempferol together with derivatives of each, (quercetin‐ O ‐glucoside, quercetin‐ O ‐xyloside), as well as kaempferol and two of its derivatives (kaempferol‐ O ‐glucoside, kaempferol‐ O ‐pentoside), respectively. Kaempferol‐ O ‐pentoside was proven by the molecular docking study as a strong inhibitor of α‐amylase and acetylcholinesterase, with binding affinities of −9.3 and −10.9 kcal/mol, respectively. As for its drug‐like properties, they were confirmed by toxicological and pharmacokinetic analyses. Hence, the present research work provides new insights into not only the promising potential use of ethyl acetate extract of R. tripartitum leaves not only for treating various diseases, but also for applications in the food processing industry.