IN-SILICO EXPERIMENTAL VALIDATION OF RUTIN FROM CITRUS MEDICA L. FOR ANTIOXIDANT AND ANTI-INFLAMMATORY POTENTIAL

Oxidative stress and chronic inflammation play a pivotal role in the pathogenesis of several non-communicable disorders, including neurodegenerative, metabolic, and cardiovascular diseases. Plant-derived flavonoids have gained increasing scientific attention owing to their antioxidant and anti-inflammatory properties. Citrus medica L. (citron), an ethnomedicinal citrus fruit, is recognized as a valuable source of bioactive phytochemicals, particularly rutin. The present investigation aimed to evaluate the nutritional composition, antioxidant potential, and prospective anti-inflammatory activity of Citrus medica flesh juice through integrated experimental and computational approaches. Proximate composition analysis was performed using standard AOAC protocols, while antioxidant activity was determined by the Ferric Reducing Antioxidant Power (FRAP) assay. Rutin content was quantified spectrophotometrically using a rutin calibration standard. Molecular docking analysis was carried out to evaluate the interaction of rutin with Toll-like receptor 4 (TLR-4), a key regulator of inflammatory signaling pathways. In addition, pharmacokinetic and drug-likeness properties of rutin were predicted using the SwissADME web platform. The proximate analysis demonstrated that Citrus medica L flesh juice possessed high moisture content (80%), along with appreciable levels of ash (8.54%), carbohydrates (8.92 g/100 g), protein (2.10 g/100 g), and crude fiber (1.57 g/100 g). The FRAP assay revealed substantial antioxidant activity with a reducing capacity of 759 mg/kg ascorbic acid equivalents, indicating significant electron-donating potential. Spectrophotometric quantification confirmed the presence of rutin at 1221 mg/kg fresh weight. Molecular docking studies demonstrated favorable interaction between rutin and TLR-4, exhibiting a binding affinity of −9.42 kcal/mol with multiple stabilizing hydrogen bond interactions involving active-site amino acid residues. SwissADME prediction indicated acceptable drug-likeness characteristics of rutin, although limitations related to gastrointestinal absorption and blood–brain barrier permeability were observed. Collectively, the findings suggest that Citrus medica flesh juice represents a promising natural source of antioxidant flavonoids with potential anti-inflammatory relevance. Nevertheless, further mechanistic investigations employing in-vitro and in-vivo experimental models are warranted to substantiate its pharmacological efficacy and translational applicability in nutraceutical and functional food development.