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.