Separation of triacylglycerols and free fatty acids in microalgal lipids by solid-phase extraction for separate fatty acid profiling analysis by gas chromatography

Paik, Man‐Jeong; Kim, Hoon; Lee, Jun Young; Brand, Jerry J.; Kim, Kyoung-Rae

doi:10.1016/j.chroma.2009.06.051

Cited by 37 publications

(27 citation statements)

References 46 publications

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“…Dry cell weight was determined after washing cells twice with H 2 O and drying overnight at 60°C. The dried lipids were transesterified with N-tertButyldimethylsilyl-N-methyltrifluoroacetamide (Sigma Aldrich) following the procedure of (Paik et al 54 ), and 2-ml samples were injected into a GC-FID (Agilent Technologies 6890 Network GC System) equipped with an Agilent HP-5 column (5% phenyl-95% methylsiloxane-product number 19091J-413) to analyse fattyacid fractions. Briefly, the following settings were used: Detector Temp ¼ 300°C, He Flow ¼ 1.0 ml min À 1 , Oven Temp ¼ 80°C for 2 min, increased at 30°C min À 1 to 200°C, increased at 2°C min À 1 to 229°C, increased at 1°C min À 1 to 232°C and increased at 50°C min À 1 to 325°C.…”

Section: Methodsmentioning

confidence: 99%

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

et al. 2014

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Economic feasibility of biosynthetic fuel and chemical production hinges upon harnessing metabolism to achieve high titre and yield. Here we report a thorough genotypic and phenotypic optimization of an oleaginous organism to create a strain with significant lipogenesis capability. Specifically, we rewire Yarrowia lipolytica's native metabolism for superior de novo lipogenesis by coupling combinatorial multiplexing of lipogenesis targets with phenotypic induction. We further complete direct conversion of lipid content into biodiesel. Tri-level metabolic control results in saturated cells containing upwards of 90% lipid content and titres exceeding 25 g l À 1 lipids, which represents a 60-fold improvement over parental strain and conditions. Through this rewiring effort, we advance fundamental understanding of lipogenesis, demonstrate non-canonical environmental and intracellular stimuli and uncouple lipogenesis from nitrogen starvation. The high titres and carbon-source independent nature of this lipogenesis in Y. lipolytica highlight the potential of this organism as a platform for efficient oleochemical production.

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Section: Methodsmentioning

confidence: 99%

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

et al. 2014

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“…Detection is often carried out by flame ionization detection, where identities of FAMEs are established by comparison of retention times to known standards [16–18], or by mass spectrometry, which offers fragmentation data for fatty acid identification [16, 18, 19]. Typically, GC analysis offers excellent separation efficiency and high-sensitivity, and provides a quantitative fatty acid profile of the lipid extract.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2011

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Biofuels 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) Electronic Supplementary MaterialSupplementary material is available for this article at 10.1007/s00216-011-5376-6 and is accessible for authorized users.

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“…In comparison to the previous analytical methods for algal sample analyses , which did not carry out method validation in algal matrix and simply applied solvent calibrators for quantitation of FAME compounds, our studies addressed the concern of matrix effect and revealed that matrix effect on analytical signals were insignificant and negligible, and the recoveries of the FAME compounds from algal matrix were high enough for accurate quantitation with the use of internal standard. Therefore, solvent calibrators instead of matrix calibrators were used in the subsequent quantitative studies.…”

Section: Resultsmentioning

confidence: 99%

Determination of fatty acid methyl esters derived from algaeScenedesmus dimorphusbiomass by GC-MS with one-step esterification of free fatty acids and transesterification of glycerolipids

Avula

Belovich

2017

J. Sep. Sci.

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Algae can synthesize, accumulate and store large amounts of lipids in its cells, which holds immense potential as a renewable source of biodiesel. In this work, we have developed and validated a GC-MS method for quantitation of fatty acids and glycerolipids in forms of fatty acid methyl esters derived from algae biomass. Algae Scenedesmus dimorphus dry mass was pulverized by mortar and pestle, then extracted by the modified Folch method and fractionated into free fatty acids and glycerolipids on aminopropyl solid-phase extraction cartridges. Fatty acid methyl esters were produced by an optimized one-step esterification of fatty acids and transesterification of glycerolipids with boron trichloride/methanol. The matrix effect, recoveries and stability of fatty acids and glycerolipids in algal matrix were first evaluated by spiking stable isotopes of pentadecanoic-2,2-d acid and glyceryl tri(hexadecanoate-2,2-d ) as surrogate analytes and tridecanoic-2,2-d acid as internal standard into algal matrix prior to sample extraction. Later, the method was validated in terms of lower limits of quantitation, linear calibration ranges, intra- and inter-assay precision and accuracy using tridecanoic-2,2-d acid as internal standard. The method developed has been applied to the quantitation of fatty acid methyl esters from free fatty acid and glycerolipid fractions of algae Scenedesmus dimorphus.

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Separation of triacylglycerols and free fatty acids in microalgal lipids by solid-phase extraction for separate fatty acid profiling analysis by gas chromatography

Cited by 37 publications

References 46 publications

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

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

Determination of fatty acid methyl esters derived from algaeScenedesmus dimorphusbiomass by GC-MS with one-step esterification of free fatty acids and transesterification of glycerolipids

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