3.2. Alkaloid Extraction

The plant material used in the present study consisted of bulbs. They were cut and dried at 40 °C, then milled with a rotary blade mill (stainless steel grinder, Taurus) into a fine powder. The powder was macerated with methanol at room temperature for 3 days, changing the solvent daily (3 × 50 mL) and applying 20 min of an ultrasonic bath 4 times per day. The mash was filtered and evaporated to dryness under reduced pressure. The crude extracts were acidified with an aqueous solution of sulfuric acid (2%, v/v) to pH 2, then cleaned with diethyl ether to remove neutral material. The aqueous solution was basified with ammonium hydroxide at 25% (v/v) to pH 9–10, then finally extracted with ethyl acetate to obtain the alkaloid extract (AE).