Ultrasound-Assisted Extraction of Total Saponins from Aralia taibaiensis: Process Optimization, Phytochemical Characterization, and Mechanism of alpha-Glucosidase Inhibition

Purpose: Aralia taibaiensis, a medicinal food plant, and total saponins from its root bark extract inhibit alpha-glucosidase activity, which is associated with type 2 diabetes; however, the inhibitory mechanism is unknown. Furthermore, a green extraction technique superior to conventional hot reflux extraction (HRE) is needed for the rapid and easy extraction of A. taibaiensis total saponins (TSAT) to exploit and utilize this resource. Our aim was to develop a green extraction method for obtaining TSAT and to investigate the mechanism by which TSAT inhibits alpha-glucosidase. Materials and Methods: In this study, the ultrasound-assisted extraction (UAE) process was optimized using a Box-Behnken design, and the extraction mechanism was investigated using scanning electron microscopy (SEM). High-performance liquid chromatography (HPLC) was used for qualitative and quantitative analyses of TSAT. In vitro glycosylation assays, enzyme kinetics, fluorescence spectroscopy measurements, atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FT-IR) and molecular docking techniques were used to investigate the mechanism by which the A. taibaiensis active ingredients inhibit alpha-glucosidase. Results: The optimal parameters for the extraction yield were obtained as an ethanol concentration of 73%, ultrasound time of 34 min, ultrasound temperature of 61 degrees C and solid-liquid ratio of 16 g/mL, which were better than HRE. The SEM analysis showed that UAE effectively disrupted plant cells, thus increasing the TSAT yield. In vitro alpha-glucosidase inhibition experiments showed that both TSAT and its active ingredient, araloside A, inhibited alpha-glucosidase activity by binding to alpha-glucosidase, thereby changing the conformation and microenvironment of alpha-glucosidase to subsequently inhibit enzyme activity. Conclusion: The optimal extraction conditions identified here established a basis for future scale-up of ultrasound extraction parameters with the potential for obtaining maximum yields. In vitro enzyme inhibition experiments investigated the mechanism of the TSAT interaction with alpha-glucosidase and further explored whether araloside A may be the main contributor to the good inhibition of alpha-glucosidase activity by TSAT.