Abstract
The interest in the physiological roles and bioactivities of plant phenols has increased over the past decades. In seaweeds, many investigations have dealt with phenolic compounds of Phaeophyceae (phlorotannins), even though little is known so far about the ecophysiological variations of their pool or their biosynthetic pathways. We describe here a simple procedure based on the use of water-organic solvent mixtures for the extraction of phlorotannins. Crude extracts are semi-purified and fractionated by separating methods based on both the polarity and the molecular size of compounds. Phenols are then quantified by the Folin-Ciocalteu method and their radical-scavenging activity is characterized using the DPPH test. All along the purification process of phenolic compounds, the efficiency of separation is assessed by 1H-NMR.
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References
Poupard P, Sanoner P, Baron A et al (2011) Characterization of procyanidin B2 oxidation products in an apple juice model solution and confirmation of their presence in apple juice by high-performance liquid chromatography coupled to electrospray ion trap mass spectrometry. J Mass Spectrom 46:1186–1197
Lattanzio V, Cardinali A, Linsalata V (2012) Introduction: plant phenolics – secondary metabolites with diverse functions. In: Cheynier V, Sarni-Manchado P, Quideau S (eds) Recent advances in polyphenol research, vol 3. Wiley, New York, pp 1–39
Stengel DB, Connan S, Popper ZA (2011) Algal chemodiversity and bioactivity: sources of natural variability and implications for commercial application. Biotechnol Adv 29:483–501
Ragan MA, Glombitza K-W (1986) Phlorotannins, brown algal polyphenols. Prog Phycol Res 4:129–241
Glombitza KW, Pauli K (2003) Fucols and phlorethols from the brown alga Scytothamnus australis Hook. et Harv. (Chnoosporaceae). Bot Mar 46:315–320
Pavia H, Toth G (2000) Influence of light and nitrogen on the phlorotannin content of the brown seaweed Ascophyllum nodosum and Fucus vesiculosus. Hydrobiologia 440:299–305
Connan S, Goulard F, Stiger V et al (2004) Phlorotannins in belt forming brown algae of a sheltered shore. Bot Mar 47:410–416
Stiger V, Deslandes E, Payri CE (2004) Phenolic contents of two brown algae, Turbinaria ornata and Sargassum mangarevense on Tahiti (French Polynesia): interspecific, ontogenic and spatio-temporal variations. Bot Mar 47:402–409
Connan S, Deslandes E, Ar Gall E (2007) Influence of nycthemeral and tidal cycles on phenol content and antioxidant capacity in three intertidal Phaeophyceae. J Exp Mar Biol Ecol 349:359–369
Zubia M, Payri C, Deslandes E (2008) Alginate, mannitol, phenolic compounds and biological activities of two range-extending brown algae, Sargassum mangarevense and Turbinaria ornata (Phaeophyta: Fucales), from Tahiti (French Polynesia). J Appl Phycol 20:1033–1043
Le Lann K, Jégou C, Stiger-Pouvreau V (2008) Effect of different conditioning treatments on total phenolic content and antioxidant activities in two Sargassacean species: comparison of the frondose Sargassum muticum (Yendo) Fensholt and the cylindrical Bifurcaria bifurcata R. Ross. Phycol Res 56:238–245
Plouguerné E, Le Lann K, Connan S et al (2006) Spatial and seasonal variation in density, reproductive status, length and phenolic content of the invasive brown macroalga Sargassum muticum (Yendo) Fensholt along the coast of Western Brittany (France). Aquat Bot 85:337–344
Ferreres F, Lopes G, Gil-Izquierdo A et al (2012) Phlorotannin extracts from Fucales characterized by HPLC-DAD-ESI-MSn: approaches to hyaluronidase inhibitory capacity and antioxidant properties. Mar Drugs 10:2766–2781
Meillisa A, Siahaan EA, Jung-Nam Park J-N et al (2013) Effect of subcritical water hydrolysate in the brown seaweed Saccharina japonica as a potential antibacterial agent on food-borne pathogens. J Appl Phycol 25:763–769
Tanniou A, Serrano Leon E, Vandanjon L et al (2013) Green improved processes to extract bioactive phenolic compounds from brown macroalgae using Sargassum muticum as model. Talanta 104:44–52
Kubanek J, Lester SE, Fenical W et al (2004) Ambiguous role of phlorotannins as chemical defences in the brown alga Fucus vesiculosus. Mar Ecol Prog Ser 277:79–93
Cérantola S, Breton F, Ar Gall E et al (2006) Co-occurrence and antioxidant activities of fucol and fucophlorethol classes of polymeric phenols in Fucus spiralis. Bot Mar 49:347–351
Zubia M, Fabre M-S, Kerjean V et al (2009) Antioxidant and antitumoural activities of some Phaeophyta from Brittany coasts. Food Chem 116:693–701
Arnold TM, Targett NM (1998) Quantifying in situ rates of phlorotannin synthesis and polymerisation in marine brown algae. J Chem Ecol 24:577–595
Breton F, Cérantola S, Ar Gall E (2011) Distribution and radical scavenging activity of phenols in Ascophyllum nodosum (Phaeophyceae). J Exp Mar Biol Ecol 399:167–172
Le Lann K, Connan S, Stiger-Pouvreau V (2012) Phenology, TPC and size-fractioning phenolics variability in temperate Sargassaceae (Phaeophyceae, Fucales) from Western Brittany: native versus introduced species. Mar Environ Res 80:1–11
Le Lann K, Ferret C, VanMee E et al (2012) Total phenolic, size-fractionated phenolics and fucoxanthin content of tropical Sargassaceae (Fucales, Phaeophyceae) from the South Pacific Ocean: spatial and specific variability. Phycol Res 60:37–50
Tierney MS, Soler-Vila A, Croft AK et al (2013) Antioxidant activity of the brown macroalga Fucus spiralis Linnaeus harvested from the West Coast of Ireland. Curr Res J Biol Sci 5:81–90
Wang T, Jónsdóttir R, Liu H et al (2012) Antioxidant capacities of phlorotannins extracted from the brown alga Fucus vesiculosus. J Agric Food Chem 60:5874–5883
Shibata T, Fujimoto K, Nagayama K et al (2002) Inhibitory activity of brown algal phlorotannins against hyaluronidase. Int J Food Sci Technol 37:703–709
Koivikko R, Eränen J, Loponen J et al (2008) Variation of phlorotannins among three populations of Fucus vesiculosus as revealed by HPLC and colorimetric quantification. J Chem Ecol 34:57–64
Audibert L, Fauchon M, Blanc N et al (2010) Phenolic compounds in the brown seaweed Ascophyllum nodosum: pool composition and radical-scavenging activities. Phytochem Anal 21:399–405
Steevensz AJ, MacKinnon SL, Hankinson R et al (2012) Profiling phlorotannins in brown macroalgae by liquid chromatography–high resolution mass spectrometry. Phytochem Anal 23:547–553
Tierney MS, Smyth TJ, Rai DK et al (2013) Enrichment of polyphenol contents and antioxidant activities of Irish brown macroalgae using food-friendly techniques based on polarity and molecular size. Food Chem 139:753–761
Parys S, Rosenbaum A, Kehraus S et al (2007) Evaluation of quantitative methods for the determination of polyphenols in algal extracts. J Nat Prod 70:1865–1870
Yotsu-Yamashita M, Kondo S, Segawa S et al (2013) Isolation and structural determination of two novel phlorotannins from the brown alga Ecklonia kurome Okamura, and their radical scavenging activities. Mar Drugs 11:165–183
Blanc N, Hauchard D, Audibert L et al (2011) Radical-scavenging capacity of phenol fractions in the brown seaweed Ascophyllum nodosum: an electrochemical approach. Talanta 84:513–518
Keyrouz R, Abasq ML, Le Bourvellec C et al (2011) Total phenolic contents, radical scavenging and cyclic voltammetry of seaweeds from Brittany. Food Chem 126:831–836
Connan S, Delisle F, Deslandes E et al (2006) Intra-thallus phlorotannin content and anti-oxidant activity in Phaeophyceae of temperate waters. Bot Mar 49:39–46
Acknowledgements
This work was supported by a grant-in-aid for scientific research (Phlorotan’ING project) from the “Groupement d’Intérêt Scientifique” Europôle Mer to V.S.P. and fellowships from the French Ministry for National Education, Higher Education and Research to M.H. and C.J. The authors are grateful to S. Cérantola, N. Kervarec, and G. Simon for support in NMR analysis and to E. Charpentier.
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Gall, E.A., Lelchat, F., Hupel, M., Jégou, C., Stiger-Pouvreau, V. (2015). Extraction and Purification of Phlorotannins from Brown Algae. In: Stengel, D., Connan, S. (eds) Natural Products From Marine Algae. Methods in Molecular Biology, vol 1308. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2684-8_7
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DOI: https://doi.org/10.1007/978-1-4939-2684-8_7
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2683-1
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