R3 rotary evaporator

Dried and powdered pods (300 g) were subjected to selective extractions with organic solvents in the following order of polarity: n-hexane, chloroform, ethyl acetate and alcohol 70% solution. The solvent: material ratio was 2:1 and the mixture was subjected to the extraction. Ultrasound-assisted extraction was performed in an ultrasonic cleaner bath (USC-1800) with a volume of 9 L, an input power of 155 W, 40 KHz of frequency, and at 30°C (± 3) and 30 min for hexane (HEX), chloroform (CLO) and acetate (ACO) extracts; and 45°C (± 3) and 30 min for aqueous ethanol extract (AE). The ultrasonic power inside de extract container was estimated to 70 W.cm-2. The extracts were concentrated with a Buchi R3 rotary evaporator (V 700 vacuum pump, V 850 vacuum controller) was used to remove the solvent at 45°C and 156 mbar, 207 mbar, 240 mbar, 240 mbar and 58 mbar pressure, respectively [22 ].

Analytical thin-layer chromatography was performed using Sorbent Technologies 250 μm glass-backed UV254 silica gel plates. The plates were first visualized by means of fluorescence upon 254 nm irradiation then using an iodine chamber and subsequently with phosphomolybdic acid with heating. Flash column chromatography was performed using Sorbent Technologies 40–63 μm, pore size 60 Å silica gel in Luknova columns on a Teledyne ISCO CombiFlash Rf with solvent systems indicated. Solvent removal was effected using a Buchi R3 rotary evaporator with a V900 diaphragm pump (~10 mmHg). Further drying of samples was conducted using a Welch vacuum pump at <0 mmHg. All isolated yields refer to material that is chromatographically (TLC or HPLC) and spectroscopically (¹H NMR) homogenous.