Fucoxanthin is the major carotenoid in oceans, found in golden-brown algae, and is considered as a bioactive molecule due to its numerous pharmaceutical and nutraceutical properties. This study aims to optimize the recovery and the purity of fucoxanthin from Tisochrysis lutea through an eco-extraction and eco-purification process, using the Centrifugal Partition Chromatography technique (CPC). CPC is based on the partition difference of fucoxanthin between two immiscible liquid phases, providing a complete sample recovery without fucoxanthin degradation, conducted at room temperature and being fast. Furthermore, CPC minimizes the amount of solvents to be effective. First, a solid-liquid US-assisted extraction was performed using dried T. lutea biomass with either acetone or ethanol. We obtained respectively 16.17 mg g−1DW and 13.54 mg g−1DW of fucoxanthin, and the specific consumption of extraction solvent was reduced by 20. Then, four experiments were conducted to optimize the organic solvent composition and the process operating conditions of the CPC. In the mobile phase, methanol used in the original method was replaced by ethanol, the amount of which has been reduced by 3 by adding H2O. The final CPC solvent system was cyclohexane:ethanol:H2O in 2:2:1 (v:v:v). Nine fractions with 100 % fucoxanthin (HPLC purity) were obtained, with 92 % of recovery yield.