Abstract
In this paper, the efficiency of pigment and fatty acid extraction from resistant algae using Scenedesmus obliquus as an example was examined. We found that adding quartz sand and solvent to freeze-dried algal material and subsequent extraction in an ultrasound bath for 90min at −4 °C resulted in excellent extraction of these compounds. This extraction method was compared with a method regularly used for extraction of fatty acids and pigments, i.e. addition of solvents to algal material with subsequent incubation. Our extraction using the ultrasound and sand method was about twice as efficient as this method for both pigments and fatty acids. The ultrasound method is simple, extracts over 90% of the different substances in one step and conserves the relationships of pigments and fatty acids. In addition, no alteration- or breakdown products were observed with the new method. Thus, this method allows accurate quantitative extraction of both pigments and fatty acids from Scenedesmus obliquus and other algae. The method was also been found to be as effective for Cryptomonas erosa (Cryptophyceae), Cyclotella meneghiniana (Bacillariophyceae), Microcystis aeruginosa (Cyanophyceae), and Staurastrum paradoxum (Chlorophyceae, Desmidiaceae) and is thus applicable to a wide spectrum of algae.
Similar content being viewed by others
References
Ackman RG, Tocher CS and McLachlan J (1968) Marine phytoplankter fatty acids. J Fish Res Bd Can 25: 1603–1620
Ahlgren G and Merino L (1991) Lipid analysis of freshwater microalgae: a method study. Arch Hydrobiol121: 295–306
AWWA-APHA (1985) Standard methods for the examination of water and waste water. 16th edition. Washington DC.
Ben-Amotz A, Tornabene TG and Thomas WH (1985) Chemical profile of selected species of microalgae with emphasis on lipids. J Phycol 21: 72–81
Bisalputra T and Weier TE (1963) The cell wall of Scenedesmus quadricauda. Am J Bot 50: 1011–1019
Bligh EG and Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917
Boersma M and Vijverberg J (1995) Synergistic effects of different food species on life-history traits of Daphnia galeata. Hydrobiologia 307: 109–115
Bowles ND, Paerl HW and Tucker J (1985) Effective solvents and extraction periods employed in phytoplankton carotenoid and chlorophyll determinations. Can J Fish Aquat Sci 42: 1127–1131
Cartens M, Grima EM, Medina AR, Gimenez AC and Gonzalez JI (1996) Eicosapentaenoic acid(20:5n-3) from the marine microalga Phaeodactylum tricornutum. J Am Oil Chem Soc 73: 1025–1031
Chen IS, Shen CSJ and Sheppard AJ (1981) Comparison of methylene chloride and chloroform for the extraction of fats from food products. J Am Oil Chem Soc 58: 599–601
Christie WW (1982) Lipid analysisq: Pergamon, Oxford
Deventer B and Heckman CW(1996) Effects of prolonged darkness on the relative pigment content of cultured diatoms and green algae. Aquatic Sci58: 241–252
Drevon CA, Baksaas I and Krokan HE (eds.) (1993) Omega-3 Fatty Acids: metabolism and Biological Effects. Birkhäuser, Basel
Fawley MW (1991) Disjunct distribution of the xanthophyll loroxanthin in the green algae (Chlorophyta). J Phycol 27: 544–548
Folch J, Lees Mand Sloane-Stanley GH (1956) A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226: 497–509
Francis GW, Knutsen G and Lien T (1973) Loroxanthin from Chlamydomonas reinhardii. Acta Chemica Scandinavica 27: 3599–3600
Jespersen AM and Christoffersen K (1987) Measurements of chlorophyll-a from phytoplankton using ethanol as extraction solvent. Arch Hydrobiol 109: 445–454
Kattner G and Fricke HSG (1986) Simple gas-liquid chromatographic method for the simultaneous determination of fatty acids and alcohols in wax esters of marine organisms. J Chrom 361: 263–268
Lürling M and van Donk E (1997) Morphological changes in Scenedesmus induced by infochemicals released in situ from zooplankton grazers. Limnol Oceanogr 42: 783–788
Mouget JL, Legendre L and De La Noüe J (1993) Initial and total activity of Rubisco in Scenedesmus ecornis: is the french press a suitable tool for enzyme extraction? Plant Cell Physiol 34: 281–288
Napolitano GE (1994) The relationship of lipids with light and chlorophyll measurements in freshwater algae and periphyton. J Phycol 30: 943–950
Parrish CC (1999) Determination of lipid, lipid classes, and fatty acids in aquatic samples. In: Arts MT and Wainmain BC (eds), Lipids in Freshwater Ecosystems. Springer-Verlag, New York
Parrish CC and Wangersky PJ (1987) Particulate and dissolved lipid classes in cultures of Phaeodactylum tricornutum grown in cage culture turbidostats with a range of nitrogen supply rates. Mar Ecol Progr Ser 35: 119–128
Rai H, Arts MT, Wainman BC, Dockal N and Krambeck HJ (1997) Lipid production in natural phytoplankton communities in a small freshwater Baltic lake, Lake Schöhsee, Germany. Freshwat Biol 38: 581–590
Strain H and SvecWA (1966) Extraction, separation, estimation and isolation of chlorophylls. In: Vernon LP and Seely GR (eds.) The Chlorophylls.
Trainor FR (1995) The sequence of ecomorph formation in a phenotypicly plastic, multispined Scenedesmus species (Chlorophyceae). Arch Hydrobiol 133: 161–171
van Donk E, Lürling M, Hessen DO and Lokhorst GM (1997) Altered cell wall morphology in nutrient-deficient phytoplankton and its impact on grazers. Limnol Oceanogr 42: 357–364
Villerius L, Wiltshire KH, Gieskes W and Paterson DM (1996) The use of a binary reversed phase, non-endcapped C30 method for the separation of algal pigments. International Carotenoid Symposium of IUPAC, Leiden, The Netherlands.
Whyte JNC (1988) Fatty acid profiles from direct methanolysis of lipids in tissue of cultured species. Aquaculture 75: 193–203
Wiltshire KH, Harsdorf S, Smidt B, Blocker g, Reuter R and Schroeder F (1998) The determination of algal biomass (as chlorophyll) in suspended matter from the Elbe estuary and the German Bight: a comparison of high-performance liquid chromatography, delayed fluorescence and prompt fluorescence methods. J Exp Marine Biol Ecol 222: 113–131
Wiltshire KH and Lampert W (1999) Urea excretion by Daphnia: A colony-inducing factor in Scenedesmus? Limnol Oceanogr 44: 1894–1903
Wood LW(1985) Chloroform-methanol extraction of chlorophyll a. Can J Fish Aquat Sci 42: 38–43
Wright SW, Jeffrey SW and Mantoura RFC (1997) Evaluation of methods and solvents for pigment extraction. In: Jeffrey SW, Mantoura RFC, and Wright SW (Eds) Phytoplankton Pigments in Oceanography: Guidelines to Modern Methods. UNESCO, Paris
Zachleder V, Wittenburg E and Abarzua S (1986) Factors controlling the inhibitory effects of 3,4-benzo(a)pyrene on the chlorococcal alga Scenedesmus quadricauda. Arch Hydrobiol Suppl 73: 281–296
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wiltshire, K.H., Boersma, M., Möller, A. et al. Extraction of pigments and fatty acids from the green alga Scenedesmus obliquus (Chlorophyceae). Aquatic Ecology 34, 119–126 (2000). https://doi.org/10.1023/A:1009911418606
Issue Date:
DOI: https://doi.org/10.1023/A:1009911418606