Natural Deep Eutectic Solvent based Ultrasound-assisted extraction: A green approach for extraction of sulfhydryl and mimosine from Leucaena leucocephala (Lam) de Wit seeds

Leucaena leucocephala (Lam.) de Wit seeds, also known as river tamarind, contain sulfhydryl compounds that exhibit antioxidant effects. However, these seeds also possess a toxic effect from mimosine. In this study, the river tamarind seeds were extracted using a natural deep eutectic solvent (NADES) based UAE. Among six NADES compositions screened, choline chloride-glycerol (ChCl-Gly) and choline chloride-sucrose (ChCl-Suc) were selected to be further optimized using a Box-Behnken Design in the RSM. The optimization of total sulfhydryl content was performed in 17 runs using three variables, namely water content in NADES (39%, 41%, and 43%), extraction time (5, 10, and 15 min), and the liquid-solid ratio (3, 5, and 7 mL/g). The highest concentration of sulfhydryls was obtained from ChCl-Gly-UAE (0.89 mg/g sample) under the conditions of a water content in NADES of 41% (v/v) and a liquid-solid ratio of 3 mL/g for 15 min, followed by that of from ChCl-Suc-UAE extract under the conditions of water content in NADES of 43% (v/v) and the liquid-solid ratio of 3 mL/g for 10 min with total sulfhydryl level was 0.67 mg/g sample. The maceration method using 30% ethanol resulted in the lowest level of sulfhydryls with a value of 0.52 mg/g. The mimosine compounds obtained in the NADES-based UAE (ChCl-Suc and ChCl-Gly) extracts were 4.95 and 7.67 mg/g, respectively, while 12.56 mg/g in the 30% ethanol-maceration extract. The surface morphology of L. leucocephala seed before and after extraction was analyzed using scanning electron microscopy. Therefore, it can be concluded that the use of ChCl-Suc and ChCl-Gly in NADES-based UAE is more selective in attracting sulfhydryl compounds than that of 30% ethanol-maceration extraction.