Natural Deep Eutectic Solvents for the Green Extraction of Betulin from Birch Bark: Chemical Characterization and Evaluation of Antioxidant Activity

Natural deep eutectic solvents (NADESs) have emerged as promising green alternatives to conventional solvents for the extraction of bioactive compounds from plant materials. In this study, eight natural deep eutectic solvents were synthesized and evaluated for their efficiency in extracting betulin from birch bark. Extraction yield was assessed using high-performance liquid chromatography with ultraviolet detection. Among the tested systems, N3 (choline chloride and urea in a 1:1 molar) and N4 (choline chloride and fructose in a 1:1 molar) were the most effective, yielding 101.26 ± 0.03 and 243.32 ± 0.26 mg betulin per gram of dry extract, respectively. Fourier transform infrared spectroscopy analysis confirmed the structural similarity of the N4 extract to pure betulin. In addition to increased extraction performance, the N4 extract demonstrated the greatest antioxidant activity (DPPH (1,1-diphenyl-2-picrylhydrazyl): 63% and ABTS (2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)): 97% inhibition) and total phenolic content (12.12 mg GAE/g extract), and betulin yield was strongly correlated with total phenolic content (TPC) and antioxidant activity (FRAP (ferric ion reducing antioxidant power), DPPH, and ABTS), indicating the preservation of bioactivity. These findings underscore the potential of NADESs as sustainable solvents for the extraction of bioactive compounds from birch bark, supporting greener extraction technologies for biomass valorization and natural product processing.