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Total Fatty Acid Analysis of Human Blood Samples in One Minute by High-Resolution Mass Spectrometry

Department of Biochemistry and Molecular Biology, Villum Center for Bioanalytical Sciences, University of Southern Denmark, 5230 Odense, Denmark
Institute of Clinical Chemistry and Laboratory Medicine, Regensburg University Hospital, 93053 Regensburg, Germany
Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford and Oxford NIHR Biomedical Research Centre, Churchill Hospital, Oxford OX3 7LE, UK
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Biomolecules 2019, 9(1), 7;
Received: 24 November 2018 / Revised: 13 December 2018 / Accepted: 17 December 2018 / Published: 27 December 2018
(This article belongs to the Special Issue Lipidomics)
PDF [2370 KB, uploaded 27 December 2018]
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Total fatty acid analysis is a routine method in many areas, including lipotyping of individuals in personalized medicine, analysis of foodstuffs, and optimization of oil production in biotechnology. This analysis is commonly done by converting fatty acyl (FA) chains of intact lipids into FA methyl esters (FAMEs) and monitoring these by gas-chromatography (GC)-based methods, typically requiring at least 15 min of analysis per sample. Here, we describe a novel method that supports fast, precise and accurate absolute quantification of total FA levels in human plasma and serum samples. The method uses acid-catalyzed transesterification with 18O-enriched H2O (i.e., H218O) to convert FA chains into 18O-labeled free fatty acids. The resulting “mass-tagged” FA analytes can be specifically monitored with improved signal-to-background by 1 min of high resolution Fourier transform mass spectrometry (FTMS) on an Orbitrap-based mass spectrometer. By benchmarking to National Institute of Standards and Technology (NIST) certified standard reference materials we show that the performance of our method is comparable, and at times superior, to that of gold-standard GC-based methods. In addition, we demonstrate that the method supports the accurate quantification of FA differences in samples obtained in dietary intervention studies and also affords specific monitoring of ingested stable isotope-labeled fatty acids (13C16-palmitate) in normoinsulinemic and hyperinsulinemic human subjects. Overall, our novel high-throughput method is generic and suitable for many application areas, spanning basic research to personalized medicine, and is particularly useful for laboratories equipped with high resolution mass spectrometers, but lacking access to GC-based instrumentation. View Full-Text
Keywords: total fatty acid analysis; human blood plasma; shotgun lipidomics; high resolution mass spectrometry; Orbitrap total fatty acid analysis; human blood plasma; shotgun lipidomics; high resolution mass spectrometry; Orbitrap

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Gallego, S.F.; Hermansson, M.; Liebisch, G.; Hodson, L.; Ejsing, C.S. Total Fatty Acid Analysis of Human Blood Samples in One Minute by High-Resolution Mass Spectrometry. Biomolecules 2019, 9, 7.

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