Antioxidant Activity, Bioavailability and Toxicity Evaluation, Density Functional Theory, Molecular Dynamics, and Molecular Simulation of Neotinea Tridentata Flower Extract

Abstract

This study aimed to investigate both the experimental (antioxidant activity, phytochemical content, and chemical profile) and computational analyses (molecular docking, DFT, toxicity profiling, and molecular dynamics) of the flower extract of Neotinea tridentata. The flowers of N. tridentata were extracted with methanol, and the antioxidant activity of the resulting extract was evaluated. Total phenolic, flavonoid, flavonol, and tannin contents were quantified using spectrophotometric methods. Additionally, the chemical profile was characterized by GC-MS; advanced computational analyses, including Absorption, Distribution, Metabolism, Excretion, And Toxicity (ADMET) predictions (SwissADME), Density functional theory (DFT) calculations (Gaussian), molecular docking (AutoDock Vina), and molecular dynamics simulations (GROMACS), were performed using the respective software programs. The methanol extract of N. tridentata exhibited strong antioxidant activity (DPPH IC₅₀: 52.05 ± 3.91 µg/mL), outperforming BHT (IC₅₀: 230 ± 10 µg/mL). The total phenolic, flavonoid, flavonol, and tannin contents were found to be 23.65 ± 2.17 mg GAE/g, 14.44 ± 0.13 mg QE/g, 25.45 ± 1.46 mg QE/g, and 2.08 ± 0.03 mg GAE/g, respectively. Gas chromatography-mass spectrometry (GC-MS) analysis revealed 18 compounds, with butanoic acid, 2-chloro-3-oxo-, methyl ester (20.39%) and xanthosine (10.20%) as major constituents. Fatty acid profiling identified cis-8,11,14-eicosatrienoic acid (20.80%) and palmitic acid (16.37%) as predominant. Molecular docking studies showed high binding affinities of key compounds, such as − 8.4 kcal/mol for tyrosinase and − 8.0 kcal/mol for xanthine oxidase. DFT analysis indicated low HOMO–LUMO gaps (e.g., 3.26 eV), suggesting high chemical reactivity and antioxidant potential. ADMET predictions indicated favorable oral bioavailability and low toxicity for major constituents. Molecular dynamics simulations (100 ns) confirmed the stability of ligand–protein complexes with RMSD values below 2.5 Å. These findings suggest that N. tridentata, with its strong antioxidant capacity and favorable drug-likeness profiles obtained through computational approaches, represents a promising candidate for further pharmacological development. © The Author(s) under exclusive licence to Society for Plant Research 2025.