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Isolation of astaxanthin monoesters from the microalgae Haematococcus pluvialis by high performance countercurrent chromatography (HPCCC) combined with high performance liquid chromatography (HPLC)
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SYSNO ASEP 0525383 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Isolation of astaxanthin monoesters from the microalgae Haematococcus pluvialis by high performance countercurrent chromatography (HPCCC) combined with high performance liquid chromatography (HPLC) Author(s) Fábryová, Tereza (MBU-M) ORCID
Tůmová, Lenka (MBU-M)
da Silva, D.C. (PT)
Pereira, D.M. (PT)
Andrade, P.B. (PT)
Valentao, P. (PT)
Hrouzek, Pavel (MBU-M) ORCID
Kopecký, Jiří (MBU-M) ORCID
Cheel, José (MBU-M) RID, ORCIDArticle number 101947 Source Title Algal Research-Biomass Biofuels and Bioproducts. - : Elsevier - ISSN 2211-9264
Roč. 49, AUG 2020 (2020)Number of pages 11 s. Language eng - English Country NL - Netherlands Keywords antioxidative properties ; mass-spectrometry ; direct extraction Subject RIV EE - Microbiology, Virology OECD category Microbiology R&D Projects LO1416 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) TJ01000013 GA TA ČR - Technology Agency of the Czech Republic (TA ČR) Method of publishing Limited access Institutional support MBU-M - RVO:61388971 UT WOS 000540736500003 EID SCOPUS 85085331841 DOI 10.1016/j.algal.2020.101947 Annotation Astaxanthin is a potent natural antioxidant with beneficial bioactivities demonstrated primarily for its free (nonesterified) form. However, its natural producer, the microalgae Haematococcus pluvialis synthesizes astaxanthin mostly in ester forms which have been little valorized so far. Hence, to contribute to the commercial use of astaxanthin esters, a scalable and efficient isolation technology is required. In this study, five astaxanthin monoesters were isolated from H. pluvialis using high performance countercurrent chromatography (HPCCC), where the lower phase of a biphasic solvent system (n-heptane:acetonitrile, ratio 5:5, v/v) was used as a mobile phase. Around 200 mg of biomass extract was subjected to the HPCCC leading to a separation of the target astaxanthin esters. To further increase the process productivity, a multi-injection HPCCC method was developed by combining two elution modes (reverse phase and co-current). In co-current elution mode, both the mobile and stationary phases were pumped simultaneously at flow rates of 3 and 1 mL/min respectively, so that the stationary phase that gets lost during each separation cycle is replenished. In total, five injections of samples (200 mg of extract, each) were achieved. Final purification with high performance liquid chromatography (HPLC) afforded five astaxanthin derivatives esterified with α-linolenic acid (1, 4 mg), linoleic acid (2, 8 mg), palmitic acid (3, 8 mg), oleic acid (4, 12 mg) and stearic acid (5, 1 mg) with purities of 98%, as determined by HPLC analysis. Only compound 4 exhibited a cytotoxic effect against human gastric cancer cells. The present study shows a useful approach for obtaining individual astaxanthin esters from H. pluvialis.
Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2021 Electronic address https://www.sciencedirect.com/science/article/pii/S2211926419312688?via%3Dihub
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