Comparison of lipid extraction methods for the microalgae Acutodesmus obliquus
Keywords:
Acutodesmus obliquus, lipid extraction, trans-esterification, Red Nile (RN)Abstract
Microalgae are widely used in the pharmaceutical and energy industries, therefore the conditions for their cultivation and extraction methods play an important role in the profiling and acquisition of lipids. The efficiency of lipid extraction from microalgae has attracted great interest from industry because of the wide variety of lipids and amounts that can be obtained. Acutodesmus obliquus (Scenedesmus obliquus UTEX 393) was used in this study. It was cultivated in Bold 3N medium modified with 75% nitrogen at 25°C, pH 6.8, 125 r/min and a photoperiod of 18/6 h and illuminated with white light provided by a Light-Emitting Diode Surface Mount Device extensions (LED SMD) with an intensity of 1200 µE/(m2•s). The cells were stained with the Red Nile (RN) technique to indicate lipid production. Four extraction methods were compared, classical, microwave (MW), Soxhlet, and ultrasound (US), using the same solvent proportions (hexane: chloroform: methanol=1:2:3). All samples were analyzed with Fourier Transform Infrared Spectroscopy (FTIR) and Gas Chromatography coupled to Mass Spectrometry (GC-MS). The results showed: 1) lipid production detected by RN was consistent with microalgal growth; 2) the MW technique was the best extraction method, according to the statistical analysis through Randomized Complete Block (RCB) design and performance of 4.6%; and 3) the presence of saturated and unsaturated acids was indicated by FTIR spectra. GC-MS was able to identify palmitic and linoleic acids as the likely major constituents of the sample. Keywords: Acutodesmus obliquus, lipid extraction, trans-esterification, Red Nile (RN) DOI: 10.25165/j.ijabe.20181105.3748 Citation: Hurtado D X, Garzón-Castro C L, Cortés-Romero J, Tello E. Comparison of lipid extraction methods for the microalgae Acutodesmus obliquus. Int J Agric & Biol Eng, 2018; 11(5): 211–217.References
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[3] Chisti Y. Biodiesel from microalgae beats bioethanol. Trends Biotechnol, 2008; 26: 126–131. doi: 10.1016/j.tibtech.2007.12.002.
[4] Bishop W M, Zubeck H M. Evaluation of microalgae for use as nutraceuticals and nutritional supplements. J Nutr Food Sci, 2012; 2. doi: 10.4172/2155-9600.1000147.
[5] Koller M, Muhr A, Braunegg G. Microalgae as versatile cellular factories for valued products. Algal Res, 2014; 6: 52–63. doi: 10.1016/j.algal.2014.09.002.
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[7] Beuckels A, Smolders E, Muylaert K. Nitrogen availability influences phosphorus removal in microalgae-based wastewater treatment. Water Res, 2015; 77: 98–106. doi: 10.1016/j.watres.2015.03.018.
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[10] Parniakov O, Barba F J, Grimi N, Marchal L, Jubeau S, Lebovka N, et al. Pulsed electric field assisted extraction of nutritionally valuable compounds from microalgae Nannochloropsis spp. using the binary mixture of organic solvents and water. Innov Food Sci Emerg Technol, 2015; 27: 79–85. doi: 10.1016/j.ifset.2014.11.002.
[11] Viswanath B, Matanda T, White S, Bux F. The microalgae – A future source of biodiesel. Microalgae-A Futur Source Biodiesel Dyn Biochem Process Biotechnol Mol Biol, 2010.
[12] Escorsim A M, da Rocha G, Vargas J V C, Mariano A B, Ramos L P, Corazza M L, et al. Extraction of Acutodesmus obliquus lipids using a mixture of ethanol and hexane as solvent. Biomass and Bioenergy, 2018; 108: 470–478. doi: 10.1016/j.biombioe.2017.10.035.
[13] Remmers I M, Hidalgo-Ulloa A, Brandt B P, Evers W A C, Wijffels R H, Lamers P P. Continuous versus batch production of lipids in the microalgae Acutodesmus obliquus. Bioresour Technol, 2017; 244: 1384–1392. doi: 10.1016/j.biortech.2017.04.093.
[14] The University of Texas at Austin. UTEX culture collection of algae at The University of Texas at Austin. UTEX 393, 2016. https://utex.org/products/utex-0393. Accessed on [2016-5-12].
[15] Li X, Hu H, Zhang Y. Growth and lipid accumulation properties of a freshwater microalga Scenedesmus sp. under different cultivation temperature. Bioresour Technol, 2011; 102: 3098–3102. doi: 10.1016/j.biortech.2010.10.055.
[16] The University of Texas at Austin. UTEX the culture collection of algae, 2014. http://web.biosci.utexas.edu/utex/mediaDetail.aspx?mediaID=55. Accessed on [2016-05-12].
[17] Hodaifa G, Martínez M E, Sánchez S. Influence of pH on the culture of Scenedesmus obliquus in olive-mill wastewater. Biotechnol Bioprocess Eng, 2010; 14: 854–860. doi: 10.1007/s12257-009-0119-7.
[18] Shen X F, Liu J J, Chu F F, Lam P K S, Zeng R J. Enhancement of FAME productivity of Scenedesmus obliquus by combining nitrogen deficiency with sufficient phosphorus supply in heterotrophic cultivation. Appl Energy, 2015; 158: 348–354. doi: 10.1016/j.apenergy.2015.08.057.
[19] Cooksey K E, Guckert J B, Williams S A, Callis P R. Fluorometric determination of the neutral lipid content of microalgal cells using Nile Red. J Microbiol Methods, 1987; 6(6): 333–345. doi: 10.1016/0167-7012(87)90019-4.
[20] Isleten-Hosoglu M, Gultepe I, Elibol M. Optimization of carbon and nitrogen sources for biomass and lipid production by Chlorella saccharophila under heterotrophic conditions and development of Nile red fluorescence based method for quantification of its neutral lipid content. Biochem Eng J, 2012; 61(4): 11-19. doi: 10.1016/j.bej.2011.12.001.
[21] Gao S, Takemura S, Ting CY, Huang S, Lu Z, Luan H, et al. The Neural Substrate of Spectral Preference in Drosophila. Neuron, 2008; 60(2): 328-342. doi: 10.1016/j.neuron.2008.08.010.
[22] Rumin J, Bonnefond H, Saint-Jean B, Rouxel C, Sciandra A, Bernard O, et al. The use of fluorescent Nile red and BODIPY for lipid measurement in microalgae. Biotechnol Biofuels, 2015; 8(1): 42. doi: 10.1186/s13068-015- 0220-4.
[23] Ott R L, Longnecker M. Statistical methods and data analysis. Seventh. Boston, 2016.
[24] Balasubramanian S, Allen J D, Kanitkar A, Boldor D. Oil extraction from Scenedesmus obliquus using a continuous microwave system--design, optimization, and quality characterization. Bioresour Technol, 2011; 102: 3396–3403. doi: 10.1016/j.biortech.2010.09.119.
[25] Florentino de Souza Silva A P, Costa M C, Colzi Lopes A, Fares Abdala Neto E, Carrhá Leitão R, Mota C R, et al. Comparison of pretreatment methods for total lipids extraction from mixed microalgae. Renew Energy, 2014; 63: 762–766. doi: 10.1016/j.renene.2013.10.038.
[26] Prabakaran P, Ravindran A D. A comparative study on effective cell disruption methods for lipid extraction from microalgae. Lett Appl Microbiol, 2011; 53: 150–154. doi: 10.1111/j.1472-765X.2011.03082.x.
[27] Koberg M, Cohen M, Ben-Amotz A, Gedanken A. Bio-diesel production directly from the microalgae biomass of Nannochloropsis by microwave and ultrasound radiation. Bioresour Technol, 2011; 102(5): 4265-4269. doi: 10.1016/j.biortech.2010.12.004.
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2018-09-29
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Hurtado, D. X., Garzón-Castro, C. L., Cortes-Romero, J., & Tello, E. (2018). Comparison of lipid extraction methods for the microalgae Acutodesmus obliquus. International Journal of Agricultural and Biological Engineering, 11(5), 211–217. Retrieved from https://ijabe.migration.pkpps03.publicknowledgeproject.org/index.php/ijabe/article/view/3748
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