Optimization of an Analytical Procedure for Extraction of Lipids from Microalgae
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An optimized procedure for extraction of total and non-polar lipids from microalgae is proposed. The effects of solvent, pretreatment (lyophilization, inactivation of lipases, and addition of antioxidants) and cell-disruption (liquid nitrogen, sonication, and bead beating) on total lipid content, lipid class, and fatty acid composition were examined. Chloroform–methanol 1:1 was shown to be the best solvent mixture for extraction of total lipids from microalgae. When performing this extraction, lyophilized algae can be used, no pretreatment with isopropanol to inactivate the lipases is needed and addition of antioxidants is not necessary. Furthermore, cell-disruption is not essential, although in that case two extractions must be performed in series to ensure that, irrespective of the microalgal species, all lipids are extracted. Determination of non-polar lipid content should be performed by separation of the total lipid extract on an SPE column. Extraction using petroleum ether is only appropriate when a bead beater is used for pretreatment.
KeywordsAlgae Lipid extraction Solvent mixture Pretreatment Cell-disruption Fat content
The research presented in this paper was financially supported by the Institute for the Promotion of Innovation by Science and Technology—Strategic Basic Research (IWT-SBO) project Sunlight and K.U.Leuven Kulak. We acknowledge IS-X (Interscience, Louvain-la-Neuve, Belgium) for the use of the GC–MS at their demolab.
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