Utilizing HPLC-DAD, multivariate analysis, molecular docking, and in vitro validation methods to investigate phenolic compounds with potential antidiabetic properties in Fraxinus mandshurica

Abstract

This comprehensive study utilized response surface methodology (RSM) to optimize extraction conditions for total flavonoids and phenols from F. mandshurica. Optimal conditions for flavonoids were 75 % ethanol, 63 °C, and 84 min, yielding 14.98 ± 0.13 mg/g. For phenols, it was 70 % ethanol, 66 °C, and 87 min, resulting in 26.04 ± 0.22 mg/g. UPLC-Q-TOF-MS/MS identified 68 constituents in F. mandshurica, categorized into groups. HPLC-DAD established a method for quantitative analysis of 12 target compounds. Geographical distribution analysis highlighted the northeast region as the main production area. Fingerprinting of 30 samples and multivariate analysis identified five active compounds. Hierarchical cluster analysis (HCA) showed differences in active compounds based on origin. In vitro antidiabetic evaluation demonstrated α-glucosidase inhibitory effects. Molecular docking studies were conducted between 12 active compounds and the antidiabetic-related protein 8A27. Esculetin had the highest binding affinity. The study examined extraction methods, analyzed samples, and explored antidiabetic effects, providing data support and insights for F. mandshurica as a natural industrial product and for extracts and active compounds as antidiabetic sources.