tAn effective high-speed counter-current chromatography (HSCCC) method was established for thepreparative isolation and purification of astaxanthin from Phaffia rhodozyma. With a two-phase sol-vent system composed of n-hexane-acetone-ethanol-water (1:1:1:1, v/v/v/v), 100 mg crude extract ofP. rhodozyma was separated to yield 20.6 mg of astaxanthin at 92.0% purity. By further one step silica gelcolumn chromatography, the purity reached 99.0%. The chemical structure of astaxanthin was confirmedby thin layer chromatography (TLC), UV spectroscopy scanning, high performance liquid chromatographywith a ZORBAX SB-C18column and a Waters Nova-pak C18column, and ESI/MS/MS.