*3.3. Preparation of Fucoxanthin*

Fucoxanthin was extracted from the brown alga, *Petalonia fascia*, cultured as described [17]. A sample (54.5 g) of frozen *Petalonia fascia* was extracted four times with 500 mL of MeOH. The combined methanol extracts were evaporated *in vacuo*. The residue was dissolved in 100 mL of 90% MeOH and then partitioned with hexane (2 × 100 mL). The 90% MeOH layer was removed to a different vessel and evaporated. The residue was resuspended in 100 mL of H2O and then extracted with EtOAc (2 × 100 mL). The EtOAc-soluble fraction (602.1 mg) was applied to a silica gel column (12 mL of Silica Gel 60, Nacalai Tesque), and the column was eluted with 170 mL of 1:1 hexane:EtOAc to give seven fractions. The fifth fraction (32.4 mg) was then separated by chromatography on ODS (10 mL of Cosmosil 75C18-OPN, Nacalai Tesque), and elution was performed with 40 mL of 85% MeOH and 30 mL of 90% MeOH. Fucoxanthin with a yield of 12.8 mg was eluted in the fraction of 90% MeOH. 
