Preparative isolation and purification of capsaicinoids from Capsicum frutescens using high-speed counter-current chromatography

Preparative high-speed counter-current chromatography (HSCCC) was successfully used for isolation and purification of capsaicinoids from an extract of Capsicum frutescens with a two-phase solvent system composed of tetrachloromethane–methanol–water at an optimized volume ratio of 4:3:2 (v/v/v). The lower phase was used as the mobile phase in the head to tail elution mode. The preparative HSCCC separation was performed on 150 mg of the crude extract yielding pure dihydrocapsaicin (33 mg), capsaicin (68 mg), and nordihydrocapsaicin (4 mg) at purities of 97.4%, 99.0% and 94.5%, respectively. The structures of the three compounds have been identified by MS, 1H NMR and 13C NMR.