Abstract
Fatty acids are a major source of structural diversity within the lipidome due to variations in their acyl chain lengths, branching, and cyclization, as well as the number, position, and stereochemistry of double bonds within their mono- and poly-unsaturated species. Here, the utility of 193 nm UltraViolet PhotoDissociation tandem mass spectrometry (UVPD-MS/MS) has been evaluated for the detailed structural characterization of a series of unsaturated fatty acid lipid species. UVPD-MS/MS of unsaturated fatty acids is shown to yield pairs of unique diagnostic product ions resulting from cleavages adjacent to their C=C double bonds, enabling unambiguous localization of the site(s) of unsaturation within these lipids. The effect of several experimental variables on the observed fragmentation behaviour and UVPD-MS/MS efficiency, including the position and number of double bonds, the effect of conjugated versus non-conjugated double bonds, the number of laser pulses, and the influence of alkali metal cations (Li, Na, K) as the ionizing adducts, has been evaluated. Importantly, the abundance of the diagnostic ions is shown to enable relative quantitation of mixtures of fatty acid isomers across a range of molar ratios. Finally, the practical application of 193 nm UVPD-MS/MS is demonstrated via characterization of changes in the ratios of fatty acid double bond positional isomers in isogenic colorectal cancer cell lines. This study therefore demonstrates the practicality of UVPD-MS/MS for the structural characterization of fatty acid isomers in lipidome analysis workflows.
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Funding
This research was supported by funding from the Australian Research Council (Grant DP190102464) to GER and OMS, and from the National Health and Medical Research Council (Grant APP1156778) to OMS and GER. OMS is a National Health and Medical Research Council Senior Research Fellow (Grant APP1136119).
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GER receives research support from Thermo Fisher Scientific. The other authors declare they have no conflict of interests.
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Published in the topical collection Current Progress in Lipidomics with guest editors Michal Holčapek, Gerhard Liebisch, and Kim Ekroos.
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Fang, M., Rustam, Y., Palmieri, M. et al. Evaluation of ultraviolet photodissociation tandem mass spectrometry for the structural assignment of unsaturated fatty acid double bond positional isomers. Anal Bioanal Chem 412, 2339–2351 (2020). https://doi.org/10.1007/s00216-020-02446-6
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DOI: https://doi.org/10.1007/s00216-020-02446-6