Isolation and purification of nootkatone from the essential oil of fruits of Alpinia oxyphylla Miquel by high-speed counter-current chromatography.

HSCCC technique in a semi-preparative-scale was successfully applied for the first time to isolation and purification of nootkatone from the essential oil of fruits of Alpinia oxyphylla Miquel. Twelve pairs of two-phase solvent systems, consisting of seven non-aqueous and five organic-aqueous solvent systems, were evaluated by HSCCC. It revealed that the separation was mainly influenced by the partition coefficient (K) of nootkatone and the separation factor (α) between nootkatone and valencene while the organic-aqueous solvent systems were more efficient than the non-aqueous systems. With the optimal two-phase solvent system composed of n-hexane-methanol-water (5:4:1, v/v) by eluting the lower phase in a head-to-tail mode, 3.1 mg of nootkatone was obtained at a purity of 92.30 % by GC-MS in one step operation from 80 mg of crude essential oil in less than 4 h. The chemical structure of nootkatone fraction was confirmed by EI-MS and (1)H NMR.