Deakin University
Browse
loke-simplemethod-2016.pdf (1.4 MB)

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

Download (1.4 MB)
Version 2 2024-06-04, 09:54
Version 1 2016-12-13, 16:11
journal contribution
posted on 2024-06-04, 09:54 authored by SNE Doomun, Stella LokeStella Loke, S O’Callaghan, Damien CallahanDamien Callahan
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.

History

Journal

Metabolites

Volume

6

Season

Special issue: Selected Paper from the Australian and New Zealand Metabolomics Conference

Article number

ARTN 42

Location

Switzerland

ISSN

2218-1989

eISSN

2218-1989

Language

English

Publication classification

C Journal article, C1 Refereed article in a scholarly journal

Copyright notice

2016, The Authors

Issue

4

Publisher

MDPI