Determination of Theobromine and Caffeine in Theobroma cacao Husk from Ethanolic Extract by GC-MS after CC Separation

Abstract

After CC separation of 21 samples from Theobroma cacao Husk was analyzed by GC-MS, from ethanolic extract at 90% was recovered a solid residue of 1 g after several days on the table of the Lab. This main residue (300 mg) was analyzed after successive separations with mixtures of CHCl 3 : MeOH (7:3) and CHCl 3 : MeOH (5:5) in CC using Sílica Gel G 60-120 Mesh. All samples were analyzed by TLC on silica gel with fluorescent indicator 254 nm on aluminum cards (layer thickness 0.2 mm) (10 × 20 cm) using CHCl 3 : MeOH: n-propanol: water (5:6:1:4) as eluent (v/v/v). The presence of Caffeine (CF) and Theobromine (TB) was confirmed after several analyses of the samples by their characteristic fragmentation pattern but nevertheless was found the presence of Theophylline.

Mass Spectrometry and Its Importance for the Analysis and Discovery of Active Molecules in Natural Products

“…The alkaloids are active ingredients whose structural feature is to have nitrogen in their structure, many of these molecules have a significant biological activity. Some alkaloids may be directly analyzed in GC/MS equipment, such as nicotine and other present in tobacco [37], caffeine and xanthine alkaloids [38,39], and others whose volatility enables its separation in gas chromatography such as tropane alkaloids [40].…”