Evaluation of anti-inflammatory activities of ethanolic extract and solvent fractions of Cleistopholis staudtii leaves

The ethanol leaf extract of Cleistopholis staudtii and its solvent fractions were evaluated for safety and bioactivity. The aim of this study was to investigate acute and chronic anti-inflammatory potential of an ethanolic leaf extract and fractions of C. staudtii in animal models. The ethanolic extract and fractions of C. staudtii leaf extract was prepared and administered orally to experimental animals used. The anti-inflammatory activity was determined by egg albumin induced paw edema in mice and formaldehyde-induced arthritis in rats. Acute oral toxicity testing revealed an LD₅₀ >5000 mg/kg with no mortality, indicating exceptional safety. Pharmacological evaluation demonstrated potent dose-dependent anti-inflammatory effects: the n-butanol fraction (500 mg/kg) showed 97.44% inhibition in acute egg albumin-induced paw edema (4 h) and 41.12% inhibition in chronic formaldehyde-induced arthritis (day 14). LC-QTOF-MS analysis of bioactive sub-fractions identified high-confidence compounds (scores >90%), including Salicin (C₁₃H₁₈O₇; precursor to aspirin; glycoside) – analgesic/anti-inflammatory; Eugenol (C₁₀H₁₂O₂; phenylpropanoid) – COX-2 inhibition & antioxidant; Kaempferol-3-O-β-diglucoside (C₂₇H₃₀O₁₆; antioxidant flavanoid) – NF-κB pathway modulation; 3,4,5-Trimethoxycinnamic acid (C₁₂H₁₄O₅; anti-inflammatory phenolic) – TNF-α suppression; and Neobavaisoflavone (C₂₀H₁₈O₄; antioxidant flavanoid) – dual COX/LOX inhibition. These results provide a scientific basis for C. staudtii's traditional use and identify its butanol fraction as a rich source of multi-target anti-inflammatory agents with clinical potential.