Isolation of Methylxantines from Cacao Beans (Theobroma cacao) by Counter-Current Chromatography (CCC)

Counter-current chromatography (CCC) was applied to the preparative separation of the methylxantines (theophylline, theobromine, and caffeine) from the methanol extract of Theobroma cacao seeds. The solvent system used was composed of hexane-ethyl acetate–methanol–water (2:3:1:1 v/v/v/v) and the extraction was monitored by high-performance liquid chromatography employing conditions for a rapid determination of the methylxantines. The mixture of methylxantines obtained were analyzed using Microsorb-MV 100-5, 3 µm (C18) column (250 mm × 4, 6 mm), and mobile phase water:methanol (75:25) in the isocratic mode at a flow-rate of 5 mL/min and detection was carried out at 275 nm.