Application of high-speed counter-current chromatography for the isolation of 9′-cis-neoxanthin from fresh spinach

Preparative HSCCC (high-speed counter-current chromatography) could be applied for the isolation of 9′-cis-neoxanthin from a crude carotenoid extract of fresh spinach leaves. The separation was performed on a Pharma-Tech Research Corp. CCC 1000 with a solvent system composed of hexane:ethanol:water at a volume ratio of 5:5:4.5 at a flow rate of 3 mL/min and at 850 rpm, using the lower phase as mobile phase. 9′-cis-neoxanthin with a purity of up to 94% could be obtained with a single HSCCC purification step of the crude carotenoid extract.