Fucoxanthin is a natural marine xanthophyll and exhibits a broad range of biological activities. In the present study, a simple and efficient two-step method was used to purify fucoxanthin from the diatom, Phaeodactylum tricornutum. The crude pigment extract of fucoxanthin was separated by silica gel column chromatography (SGCC). Then, the fucoxanthin-rich fraction was purified using a hydrophile-lipophile balance (HLB) solid-phase extraction column. The identification and quantification of fucoxanthin were determined by high-performance liquid chromatography (HPLC) and electrospray ionization mass spectrometry (ESI-MS). This two-step method can obtain 92.03% pure fucoxanthin and a 76.67% recovery rate. In addition, 1H and 13C NMR spectrums were adopted to confirm the identity of fucoxanthin. Finally, the purified fucoxanthin exhibited strong antioxidant properties in vitro with the effective concentration for 50% of maximal scavenging (EC50) of 1,1-Dihpenyl-2-picrylhydrazyl (DPPH) and 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radicals being 0.14 mg·mL-1 and 0.05 mg·mL-1, respectively.
Keywords: ESI-MS; HLB solid-phase extraction; HPLC; NMR; SGCC; antioxidant; fucoxanthin; purification.