Tetrabutylammonium bromide-based hydrophobic deep eutectic solvent for the extraction and separation of dihydromyricetin from vine tea and its inhibitory efficiency against xanthine oxidase

Abstract

In this study, deep eutectic solvent oscillation-assisted extraction (DES-OS) combined with macroporous resin adsorption and desorption technology was used to achieve the rapid green extraction and separation of the characteristic component dihydromyricetin (DMY) from vine tea. Multivariate data analysis showed that the DES system composed of tetrabutylammonium bromide (N₄₄₄Br) and pyruvic acid (molar ratio 1 : 2) had good extraction performance for DMY. The influence parameters of DES-OS were studied, and optimized by the single-factor test and response surface methodology (RSM) with Box–Behnken design (BBD). The extraction model of DMY was established and verified. The results showed that the extraction yield of DMY could reach 40.1 mg g⁻¹ under the optimal conditions (DES water contents of 71.18%, extraction time of 2.80 h, extraction temperature of 46.40 °C), which is in good agreement with the predicted value. In addition, Fourier transform infrared spectroscopy (FT-IR) was used to characterize the solvent before and after extraction. Scanning electron microscopy (SEM) results further confirmed that tetrabutylammonium bromide:pyruvate enhanced the destruction of the cell wall structure, resulting in the release of more DMY. Furthermore, different macroporous resins were selected for the separation of DMY for the DES-OS extract, and it was found that the DM301 resin had the ideal recovery performance under optimized dynamic condition. Finally, the product was found to have an inhibitory effect against xanthine oxidase (XO) as a mixed-type competitive inhibitor with IC50 values of (5.79 ± 0.22) × 10⁻⁵ mol L⁻¹. The inhibitory mechanisms of DMY on XO were explored by enzyme kinetics, spectroscopy, molecular docking and molecular dynamics analysis approaches, which provided a theoretical basis for the above inhibition assays.