Abstract
The increasing interest in the analysis of triglyceride (TG) species and the individual fatty acid (FA) composition requires expeditious and reliable quantification strategies. The utilization of flow injection analysis (FIA) coupled to quadrupole tandem mass spectrometry (MS/MS) for the simultaneous quantitation of TG and identification of FA composition facilitates the multiplexed verification of various biomarkers from small sample quantities. Enzymatic methods based on saponification and glycerol analysis are not suited for the determination of the FA distribution in TGs. This protocol proposes a procedure for the establishment of a relative quantitation method for middle- to high-abundance plasma TGs and the corresponding FA composition. Essential topics as FIA-MS/MS method development as well as sample preparation and validation strategies are described in detail.
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References
Erkkilä A, de Mello VDF, Risérus U, Laaksonen DE (2008) Dietary fatty acids and cardiovascular disease: an epidemiological approach. Prog Lipid Res 47:172–187. https://doi.org/10.1016/j.plipres.2008.01.004
Rhee EP et al (2011) Lipid profiling identifies a triacylglycerol signature of insulin resistance and improves diabetes prediction in humans. J Clin Invest 121(4):1402–1411. https://doi.org/10.1172/JCI44442
Nagy K, Sandoz L, Destaillats F, Schafer O (2013) Mapping the regioisomeric distribution of fatty acids in triacylglycerols by hybrid mass spectrometry. J Lipid Res 54(1):290–305. https://doi.org/10.1194/jlr.D031484
Baiocchi C, Medana C, Dal Bello F (2015) Analysis of regioisomers of polyunsaturated triacylglycerols in marine matrices by HPLC/HRMS. Food Chem 166:551–560. https://doi.org/10.1016/j.foodchem.2014.06.067
Han X, Gross RW (2001) Quantitative analysis and molecular species fingerprinting of triacylglyceride molecular species directly from lipid extracts of biological samples by electrospray ionization tandem mass spectrometry. Anal Biochem 295(1):88–100. https://doi.org/10.1006/abio.2001.5178
Krank J, Murphy RC, Barkley RM, Duchoslav E, McAnoy A (2007) Qualitative analysis and quantitative assessment of changes in neutral glycerol lipid molecular species within cells. In: Brown HA (ed) Methods in enzymology, vol 432. Academic Press, USA, pp 1–20. https://doi.org/10.1016/S0076-6879(07)32001-6.
Fernandez C, Sandin M, Sampaio JL, Almgren P, Narkiewicz K, Hoffmann M, Hedner T, Wahlstrand B, Simons K, Shevchenko A, James P, Melander O (2013) Plasma lipid composition and risk of developing cardiovascular disease. PLoS One 8(8):e71846
Schwudke D, Oegema J, Burton L, Entchev E, Hannich JT, Ejsing CS, Kurzchalia T, Shevchenko A (2005) Lipid profiling by multiple precursor and neutral loss scanning driven by the data-dependent acquisition. Anal Chem 78(2):585–595. https://doi.org/10.1021/ac051605m.
Hsu F-F, Turk J (2010) Electrospray ionization multiple-stage linear ion-trap mass spectrometry for structural elucidation of triacylglycerols: assignment of fatty acyl groups on the glycerol backbone and location of double bonds. J Am Soc Mass Spectrom 21(4):657–669. https://doi.org/10.1016/j.jasms.2010.01.007
Sander M, Becker S, Thiery J, Ceglarek U (2015) Simultaneous identification and quantification of triacylglycerol species in human plasma by flow-injection electrospray ionization tandem mass spectrometry. Chromatographia 78(5):435–443. https://doi.org/10.1007/s10337-014-2782-x
Meyer VR (2010) Practical high-performance liquid chromatography. John Wiley & Sons, Ltd, Chichester, UK
Leiker TJ, Barkley RM, Murphy RC (2011) Analysis of diacylglycerol molecular species in cellular lipid extracts by normal-phase LC-electrospray mass spectrometry. Int J Mass Spectrom 305:103–108. https://doi.org/10.1016/j.ijms.2010.09.008
Quehenberger O et al (2010) Lipidomics reveals a remarkable diversity of lipids in human plasma. J Lipid Res 51:3299–3305. https://doi.org/10.1194/jlr.M009449
Acknowledgment
This publication is supported by LIFE—Leipzig Research Center for Civilization Diseases—Leipzig University. LIFE is funded by means of the European Union, by the European Regional Development Fund (ERDF), and by the Free State of Saxony within the framework of the excellence initiative.
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Reinicke, M., Becker, S., Ceglarek, U. (2018). LC-MS/MS Analysis of Triglycerides in Blood-Derived Samples. In: Giera, M. (eds) Clinical Metabolomics. Methods in Molecular Biology, vol 1730. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7592-1_9
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DOI: https://doi.org/10.1007/978-1-4939-7592-1_9
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