Updated pigment composition of Tisochrysis lutea and purification of fucoxanthin using centrifugal partition chromatography coupled to flash chromatography for the chemosensitization of melanoma cells

Abstract

Tisochrysis lutea (T. lutea, ex T. Isochrysis galbana or T-Iso) is a marine haptophyte that was first isolated from Tahiti seawater. Because of its high content in lipids, this tropical species is commonly used in aquaculture to feed fishes, crustaceans and molluscs larvae. It is also a rich source of fucoxanthin with a high potential for nutraceutical, cosmetic and pharmaceutical applications. The purpose of the present study was to detail the pigment composition of T. lutea and to develop an efficient process to recover highly purified fucoxanthin. Using ultra performant liquid chromatography coupled to diode arrays and high-resolution mass spectrometry detectors (UPLC-DAD-MS/MS), we demonstrated for the first time the presence of echinenone, 3-hydroxy-echinenone and chlorophyll c2-monogalactosyldiacylglycerol [18:4/14:0] in unstressed cultures of T. lutea. The chemotaxonomic relevance of this updated pigment composition was discussed in relation to the Haptophyta phylum. A two-step purification of fucoxanthin was then optimized using centrifugal partition chromatography coupled to flash chromatography. This process allowed the efficient isolation of fucoxanthin (purity > 99%), that was further assessed as a lowtoxicity antineoplastic and chemosensitizing natural product in human chemoresistant melanoma cells. This carotenoid exerted an antiproliferative activity in A2058 melanoma cells and reversed in vitro their chemoresistance to dacarbazine, a DNA-alkylating agent clinically used for the treatment of metastatic melanoma.Please note that this is an author-produced PDF of an article accepted for publication following peer review. The definitive publisher-authenticated version is available on the publisher Web site.

Production of Fucoxanthin from Phaeodactylum tricornutum Using High Performance Countercurrent Chromatography Retaining Its FOXO3 Nuclear Translocation-Inducing Effect

“…Fucoxanthin has been mostly obtained from algal biomass using liquid-solid chromatography applying multi-step procedures [ 25 , 28 , 29 , 30 , 31 , 32 , 33 ]. Efforts have been dedicated to the production of fucoxanthin from algae using liquid-liquid chromatography namely CCC or CPC [ 50 , 51 , 52 ]; however, little has been done to improve the efficiency of the process for obtaining microalgae-sourced fucoxanthin. Unlike solid–liquid chromatography, no expensive columns are required in countercurrent chromatography [ 49 , 60 ]; thus, its use may represent a significant cost saving.…”

“…In this study, the developed separation process using a multiple-sequential injection strategy was aimed to enable the quick and large-scale isolation of fucoxanthin from the diatom P. tricornutum . Different liquid-liquid separation methods using CCC or CPC have already been reported for the separation of fucoxanthin from different algae species [ 50 , 51 , 52 ]. Two of these methods processed directly crude extracts using CPC for one microalgae species [ 52 ] and CCC for three macroalgae species [ 50 ].…”

“…Different liquid-liquid separation methods using CCC or CPC have already been reported for the separation of fucoxanthin from different algae species [ 50 , 51 , 52 ]. Two of these methods processed directly crude extracts using CPC for one microalgae species [ 52 ] and CCC for three macroalgae species [ 50 ]. A third reported method [ 51 ] involved a previous fractionation of macroalgal extract by solvent partitioning before CPC separation.…”

“…High-speed countercurrent chromatography (HSCCC) has been used for the isolation of fucoxanthin from edible brown macroalgae species such as Laminaria japonica , Undaria pinnatifida and Sargassum fusiforme [ 50 ] applying a single sample injection method. Centrifugal partition chromatography (CPC), another variant of liquid-liquid chromatography, was also used in two steps to isolate fucoxanthin from the macroalgae Eisenia bicyclis [ 51 ] and in one step followed by flash chromatography to produce fucoxanthin from the microalgae Tisochrysis lutea [ 52 ]. The present study reports, for the first time, an efficient and scalable HPCCC isolation method to obtain fucoxanthin from the diatom P. tricornutum using a multiple sequential-injection separation strategy.…”

Xanthophylls from the Sea: Algae as Source of Bioactive Carotenoids

“…In addition, in vivo studies were also performed. For example, in mice, the administration of FU suppressed tumor growth of primary effusion lymphoma, sarcomas, and osteosarcoma [ 51 , 55 , 96 ]. Due to its anti-inflammatory activity, FU is also being tested to prevent and treat inflammatory-related diseases, thanks to fucoxanthin’s strong antioxidant capacity and gut microbiota regulation [ 99 ], and its capacity to inhibit the production of nitric oxide, which is one of the determinants of inflammation in cells [ 100 ].…”