Vaccum controller v 800

MAE was accomplished with a MARS-X 1500 W (Microwave Accelerated Reaction System for Extraction and Digestion, CEM, Mathews, NC, USA) using closed Teflon extraction vessels. Briefly, samples of dried powdered leaf (300 mg) were extracted with 10 mL of solvent (water, ethanol and ethanol:water (50%, v/v)) [31 (link)]. Microwave power was fixed at 300 W and the extraction was performed at selected temperatures (72–94 °C) during 1 to 5 min, with constant medium stirring, according to Benlloch-Tinoco et al. [32 (link)], with minor modifications. After extraction, samples were filtrated through a Whatman No. 1 filter paper. The aqueous extract was frozen at −80 °C for subsequent lyophilization (Telstar, model Cryodos -80, Spain), while the alcoholic extract was evaporated at 40 °C (Vaccum Controller V-800, Büchi, Switzerland). The hydroalcoholic extract was firstly evaporated and then lyophilized to eliminate the residual water. Lastly, samples were stocked at room temperature (20 °C) and covered from light until further analysis.

D. crassifolium extracts were obtained via UAE using an ultrasonic probe processor (Sonic Vibracell, model VCX50, Newtown, CT, USA) and a probe tip (No. 630-0219) that was 13 mm in diameter, using the optimal extraction conditions determined by Silva et al. [11 (link)], namely a solid–liquid ratio of 10% (w/v) and an ultrasonic intensity of 30 W/m² for 31.11 min. Water and absolute ethanol were used as solvents. The aqueous extract was frozen at −80 °C for subsequent lyophilization (Telstar, model Cryodos-80, Barcelona, Spain), while the alcoholic one was evaporated at 40 °C (Vaccum Controller V-800, Büchi, Flawil, Switzerland). Samples were kept at room temperature until further analysis.