Phyto-Metabolomic Insights Into Ficus hispida Linn.: Antioxidant Activity, Polyphenolic Content, and Antidiabetic Marker Discovery via LC-ESI-Orbitrap-MS/MS and GC-QqQ-MS/MS.
Abstract
The wild edible fruit Ficus hispida Linn (F. hispida), traditionally used for diabetes management, remains unexplored with respect to bioactive constituents and underlying mechanisms. Therefore, this study investigated the antidiabetic potential of F. hispida fruits through comprehensive phyto-metabolomic profiling integrated with biochemical assays and in silico approaches. Fruit extracts were prepared via cold maceration and evaluated for total phenolic and flavonoid contents, antioxidant activity, and inhibition of α-amylase and α-glucosidase. Comprehensive untargeted metabolite profiling was performed using GC-QqQ-MS/MS and LC-ESI-Orbitrap-MS/MS, while targeted analysis was conducted by HPLC. Metabolite annotation and data processing were conducted using Compound Discoverer (v3.2.2.31) and NIST libraries, followed by PCA, PLS-DA, and hierarchical clustering using MetaboAnalyst 6.0. Methanol and hydroalcoholic extracts exhibited the highest phenolic (78.85 ± 0.58 and 74.68 ± 0.24 mg GAE/g) and flavonoid contents (89.03 ± 0.45 and 71.91 ± 1.96 mg NE/g, respectively), which corresponded with their stronger α-amylase inhibition (IC50: 91.11 ± 3.54 and 75.96 ± 3.48 µg/mL) and α-glucosidase inhibition (IC50: 101.44 ± 2.04 and 89.29 ± 3.03 µg/mL, respectively). GC‒MS/MS and LC‒MS/MS profiling revealed 160 and 759 putatively annotated metabolites (MSI 2), respectively. Furthermore, in silico molecular docking identified new α-amylase and α-glucosidase inhibitors, that is, desmodol (-8.324 kcal/mol) and demethylbatatasin IV (-7.062 kcal/mol), respectively. Complementarily, molecular dynamics simulation demonstrated the stable binding of both desmodol-α-amylase and demethylbatastasin IV-α-glucosidase complexes, supporting their potential therapeutic relevance as antidiabetic phytoconstituents. Overall, this study highlights the potential of integrated phyto-metabolomics and chemometrics for identifying antidiabetic markers with applications in functional foods and therapeutics.