NADES-assisted sustainable extraction of total protein content from Achyranthes bidentata seeds and their biochemical characterization.

This study evaluates Natural Deep Eutectic Solvents (NADES) as sustainable alternatives for the extraction of proteins and phytochemicals from Achyranthes bidentata seeds. Six lactic acid-based NADES formulations were compared against water and ethanol using ultrasound-assisted extraction. Total protein, total phenolic content (TPC), and total flavonoid content (TFC) were quantified. NADES significantly outperformed conventional solvents in extraction efficiency. The binary NADES N1 (lactic acid: glycerol, 1:1) achieved the highest protein concentration (318.55 ± 24.57 μg/mL). In contrast, the ternary NADES N5 (lactic acid: glycerol: choline chloride, 2:1:1) exhibited the highest efficiency for secondary metabolites, yielding a TPC of 16.33 ± 4.61 mg GAE/g and TFC of 409.62 ± 60.47 mg QE/g. These findings demonstrate the tunable nature of NADES for targeted biomolecule recovery, validating A. bidentata seeds as a promising source of functional ingredients utilizing green extraction methodologies.