Evaluation of Extraction Techniques, Chemical Composition, Antioxidant Properties, Computational Modelling, and Biological Insights of the Novel Truffle Species (Tirmania Nivea) for Enhanced Human Health Benefits

Abstract

This study investigates the chemical composition, antioxidant properties, and theoretical insights into the antioxidant activity of Libyan truffle T. nivea, using extracts from petroleum ether, chloroform, and methanol. The truffles contained 12.64% protein, 6.56% fat, 5.91% fiber, and 2.58% ash, indicating their significant nutritional profile. The methanol extract showed the highest antioxidant activity with 106 mg of total phenolics (GAE/100g) and 71 mg of flavonoids (QE/100 g), resulting in 86.30% inhibition in the DPPH• assay at 1,000 μg/mL and 96.9% inhibition in the β-carotene bleaching assay at 10 µg/mL. The chloroform extract exhibited 74.88% DPPH• inhibition at 1,000 μg/mL and 85.1% inhibition at 0.5 µg/mL in the β-carotene assay, while the petroleum ether extract showed 69.09% inhibition at 1000 μg/mL in DPPH• and lower activity in β-carotene bleaching. Theoretical calculations using Density Functional Theory (DFT) revealed that phenolic compounds such as quercetin and rosmarinic acid, which possess lower energy gaps (Eg), exhibited stronger reactivity and higher antioxidant potential. For example, quercetin (Eg = 4.034 eV) and rosmarinic acid (Eg = 3.50 eV) exhibited significant biological activity due to their smaller energy gaps, indicating higher reactivity toward free radicals. In contrast, compounds such as caffeic acid (Eg = 4.148 eV) and benzoic acid (Eg = 5.76 eV) showed larger energy gaps, leading to lower reactivity and antioxidant effects. The results also indicated that hydroxyl-rich compounds, such as pyrogallol and gallic acid, had high electron density and superior antioxidant performance. Molecular docking simulations demonstrated strong binding of phenolic compounds like rosmarinic acid (‒16.15 kcal/mol) and quercetin (‒15.14 kcal/mol) with xanthine oxidase, suggesting their potential as potent antioxidants. These findings highlight the potential of T. nivea truffles, particularly the methanol extract, as a rich source of natural antioxidants with substantial nutritional and therapeutic benefits. HIGHLIGHTS Libyan truffle T. nivea exhibits significant antioxidant potential. Methanol extract shows superior activity among tested solvents. Phenolic compounds are major contributors to antioxidant effects. Theoretical and docking analyses support their reactivity and enzyme interactions. GRAPHICAL ABSTRACT