Triacylglycerol profiling of microalgae strains for biofuel feedstock by liquid chromatography–high-resolution mass spectrometry

AbstractBiofuels from photosynthetic microalgae are quickly gaining interest as a viable carbon-neutral energy source. Typically, characterization of algal feedstock involves breaking down triacylglycerols (TAG) and other intact lipids, followed by derivatization of the fatty acids to fatty acid methyl esters prior to analysis by gas chromatography (GC). However, knowledge of the intact lipid profile could offer significant advantages for discovery stage biofuel research such as the selection of an algal strain or the optimization of growth and extraction conditions. Herein, lipid extracts from microalgae were directly analyzed by ultra-high pressure liquid chromatography–mass spectrometry (UHPLC-MS) using a benchtop Orbitrap mass spectrometer. Phospholipids, glycolipids, and TAGs were analyzed in the same chromatographic run, using a combination of accurate mass and diagnostic fragment ions for identification. Using this approach, greater than 100 unique TAGs were identified over the six algal strains studied and TAG profiles were obtained to assess their potential for biofuel applications. Under the growth conditions employed, Botryococcus braunii and Scenedesmus obliquus yielded the most comprehensive TAG profile with a high abundance of TAGs containing oleic acid. FigureOptical microscope image of Botryococcus braunii and high resolution mass spectrum of triacylglycerol 28:2/18:1/18:1 (inset)

[1]  E. Hvattum Analysis of triacylglycerols with non-aqueous reversed-phase liquid chromatography and positive ion electrospray tandem mass spectrometry. , 2001, Rapid communications in mass spectrometry : RCM.

[2]  Yu Bai,et al.  Recent advances of chromatography and mass spectrometry in lipidomics , 2011, Analytical and bioanalytical chemistry.

[3]  Y. Chisti Biodiesel from microalgae. , 2007, Biotechnology advances.

[4]  A Ruth Godfrey,et al.  Accurate mass measurement: Terminology and treatment of data , 2010, Journal of the American Society for Mass Spectrometry.

[5]  R. Wijffels,et al.  An Outlook on Microalgal Biofuels , 2010, Science.

[6]  Philip Owende,et al.  Biofuels from microalgae—A review of technologies for production, processing, and extractions of biofuels and co-products , 2010 .

[7]  Ryo Taguchi,et al.  Global analysis of triacylglycerols including oxidized molecular species by reverse-phase high resolution LC/ESI-QTOF MS/MS. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[8]  W. Byrdwell,et al.  Dual parallel liquid chromatography with dual mass spectrometry (LC2/MS2) for a total lipid analysis. , 2008, Frontiers in bioscience : a journal and virtual library.

[9]  J. Brand,et al.  Separation of triacylglycerols and free fatty acids in microalgal lipids by solid-phase extraction for separate fatty acid profiling analysis by gas chromatography. , 2009, Journal of chromatography. A.

[10]  C. Posten,et al.  Second Generation Biofuels: High-Efficiency Microalgae for Biodiesel Production , 2008, BioEnergy Research.

[11]  Rashmi,et al.  Prospects of biodiesel production from microalgae in India , 2009 .

[12]  C. Howe,et al.  Biodiesel from algae: challenges and prospects. , 2010, Current opinion in biotechnology.

[13]  A. Tchapla,et al.  Simple complementary liquid chromatography and mass spectrometry approaches for the characterization of triacylglycerols in Pinus koraiensis seed oil. , 2011, Journal of chromatography. A.

[14]  Anoop Singh,et al.  Renewable fuels from algae: an answer to debatable land based fuels. , 2011, Bioresource technology.

[15]  Eoin Fahy,et al.  LIPID MAPS online tools for lipid research , 2007, Nucleic Acids Res..

[16]  M. Mansour Reversed-phase high-performance liquid chromatography purification of methyl esters of C(16)-C(28) polyunsaturated fatty acids in microalgae, including octacosaoctaenoic acid [28:8(n-3)]. , 2005, Journal of chromatography. A.

[17]  Michal Holcapek,et al.  Triacylglycerols profiling in plant oils important in food industry, dietetics and cosmetics using high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. , 2008, Journal of chromatography. A.

[18]  Christoph Benning,et al.  Plant triacylglycerols as feedstocks for the production of biofuels. , 2008, The Plant journal : for cell and molecular biology.

[19]  R. Guillard,et al.  Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt, and Detonula confervacea (cleve) Gran. , 1962, Canadian journal of microbiology.

[20]  Albert Koulman,et al.  RAPID COMMUNICATIONS IN MASS SPECTROMETRY Rapid Commun. Mass Spectrom. 2009; 23: 1411–1418 , 2022 .

[21]  Jun Zhu,et al.  Anaerobic digested dairy manure as a nutrient supplement for cultivation of oil-rich green microalgae Chlorella sp. , 2010, Bioresource technology.

[22]  D. Bilanović,et al.  Freshwater and marine microalgae sequestering of CO2 at different C and N concentrations – Response surface methodology analysis , 2009 .

[23]  C. Lan,et al.  CO2 bio-mitigation using microalgae , 2008, Applied Microbiology and Biotechnology.

[24]  Q. Hu,et al.  Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. , 2008, The Plant journal : for cell and molecular biology.

[25]  Yeping Xiong,et al.  Measurement of phospholipids by hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry: the determination of choline containing compounds in foods. , 2011, Journal of chromatography. A.

[26]  Michal Holčapek,et al.  Lipidomic profiling of biological tissues using off-line two-dimensional high-performance liquid chromatography-mass spectrometry. , 2011, Journal of chromatography. A.

[27]  M. Suematsu,et al.  Triacylglycerol/phospholipid molecular species profiling of fatty livers and regenerated non-fatty livers in cystathionine beta-synthase-deficient mice, an animal model for homocysteinemia/homocystinuria , 2011, Analytical and bioanalytical chemistry.

[28]  Ranran Liu,et al.  Lipid profiling of rat peritoneal surface layers by online normal- and reversed-phase 2D LC QToF-MS[S] , 2010, Journal of Lipid Research.

[29]  L. Rodolfi,et al.  Microalgae for oil: Strain selection, induction of lipid synthesis and outdoor mass cultivation in a low‐cost photobioreactor , 2009, Biotechnology and bioengineering.

[30]  L. Laurens,et al.  Microalgae as biodiesel & biomass feedstocks: Review & analysis of the biochemistry, energetics & economics , 2010 .

[31]  J. Pittman,et al.  The potential of sustainable algal biofuel production using wastewater resources. , 2011, Bioresource technology.

[32]  Y. Chisti,et al.  Botryococcus braunii: A Renewable Source of Hydrocarbons and Other Chemicals , 2002, Critical reviews in biotechnology.

[33]  Yen-Hui Lin,et al.  Influence of growth phase and nutrient source on fatty acid composition of Isochrysis galbana CCMP 1324 in a batch photoreactor , 2007 .

[34]  W. Lindner,et al.  Selective enrichment of phosphatidylcholines from food and biological matrices using metal oxides as solid-phase extraction materials prior to analysis by HPLC–ESI-MS/MS , 2010, Analytical and bioanalytical chemistry.

[35]  J. Melanson,et al.  Quantitative analysis of positional isomers of triacylglycerols via electrospray ionization tandem mass spectrometry of sodiated adducts. , 2010, Rapid communications in mass spectrometry : RCM.

[36]  Michael J. Thomas,et al.  Phospholipid profiling by tandem mass spectrometry. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[37]  Qingyu Wu,et al.  Pyrolytic characteristics of heterotrophic Chlorella protothecoides for renewable bio-fuel production , 2001, Journal of Applied Phycology.

[38]  Frank David,et al.  Comprehensive blood plasma lipidomics by liquid chromatography/quadrupole time-of-flight mass spectrometry. , 2010, Journal of chromatography. A.