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A simple method for measuring carbon-13 fatty acid enrichment in the major lipid classes of microalgae using GC-MS

Elahee Doomun, Sheik Nadeem, Loke, Stella, O'Callaghan, Sean and Callahan, Damien L 2016, A simple method for measuring carbon-13 fatty acid enrichment in the major lipid classes of microalgae using GC-MS, Metabolites, vol. 6, no. 4, Special issue: Selected Paper from the Australian and New Zealand Metabolomics Conference, pp. 1-13, doi: 10.3390/metabo6040042.

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Title A simple method for measuring carbon-13 fatty acid enrichment in the major lipid classes of microalgae using GC-MS
Author(s) Elahee Doomun, Sheik Nadeem
Loke, Stella
O'Callaghan, Sean
Callahan, Damien LORCID iD for Callahan, Damien L orcid.org/0000-0002-6384-8717
Journal name Metabolites
Volume number 6
Issue number 4
Season Special issue: Selected Paper from the Australian and New Zealand Metabolomics Conference
Start page 1
End page 13
Total pages 13
Publisher MDPI
Place of publication Basel, Switzerland
Publication date 2016-12
ISSN 2218-1989
Keyword(s) 13C stable isotope measurement
chemical ionisation
gas chromatography
mass spectrometry
solid-phase extractions
algae
lipids
Summary A simple method for tracing carbon fixation and lipid synthesis in microalgae was developed using a combination of solid-phase extraction (SPE) and negative ion chemical ionisation gas chromatography mass spectrometry (NCI-GC-MS). NCI-GC-MS is an extremely sensitive technique that can produce an unfragmented molecular ion making this technique particularly useful for stable isotope enrichment studies. Derivatisation of fatty acids using pentafluorobenzyl bromide (PFBBr) allows the coupling of the high separation efficiency of GC and the measurement of unfragmented molecular ions for each of the fatty acids by single quadrupole MS. The key is that isotope spectra can be measured without interference from co-eluting fatty acids or other molecules. Pre-fractionation of lipid extracts by SPE allows the measurement of13C isotope incorporation into the three main lipid classes (phospholipids, glycolipids, neutral lipids) in microalgae thus allowing the study of complex lipid biochemistry using relatively straightforward analytical technology. The high selectivity of GC is necessary as it allows the collection of mass spectra for individual fatty acids, including cis/trans isomers, of the PFB-derivatised fatty acids. The combination of solid-phase extraction and GC-MS enables the accurate determination of13C incorporation into each lipid pool. Three solvent extraction protocols that are commonly used in lipidomics were also evaluated and are described here with regard to extraction efficiencies for lipid analysis in microalgae.
Language eng
DOI 10.3390/metabo6040042
Field of Research 030101 Analytical Spectrometry
Socio Economic Objective 970103 Expanding Knowledge in the Chemical Sciences
HERDC Research category C1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2016, The Authors
Free to Read? Yes
Use Rights Creative Commons Attribution licence
Persistent URL http://hdl.handle.net/10536/DRO/DU:30090215

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Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.