Separation and purification of astaxanthin from Phaffia rhodozyma by preparative high-speed counter-current chromatography

•Astaxanthin was obtained from Phaffia Rhodozyma using HSCCC for the first time.•The purity of astaxanthin was higher than the previously reported literature.•The separation and purification process of astaxanthin was simpler and more efficient than traditional methods.•TLC, UV spectroscopy scaning, HPLC and ESI/MS/MS were used to identify astaxanthin.

An effective high-speed counter-current chromatography (HSCCC) method was established for the preparative isolation and purification of astaxanthin from Phaffia rhodozyma. With a two-phase solvent system composed of n-hexane-acetone-ethanol-water (1:1:1:1, v/v/v/v), 100 mg crude extract of P. rhodozyma was separated to yield 20.6 mg of astaxanthin at 92.0% purity. By further one step silica gel column chromatography, the purity reached 99.0%. The chemical structure of astaxanthin was confirmed by thin layer chromatography (TLC), UV spectroscopy scanning, high performance liquid chromatography with a ZORBAX SB-C18 column and a Waters Nova-pak C18 column, and ESI/MS/MS.