Abstract
Metabolomics aims at the identification and quantification of key-end point metabolites, basically polar, in order to study changes in biochemical activities in response to pathophysiological stimuli or genetic modifications. Targeted profiling assays have enjoyed a growing popularity during the last years with LC-MS/MS as a powerful tool for development of such (semi-) quantitative methods for a large number of metabolites. Here we describe a method for absolute quantification of ca. 100 metabolites belonging to key metabolite classes such as sugars, amino acids, nucleotides, organic acids, and amines with a hydrophilic interaction liquid chromatography (HILIC) system comprised of ultra (high) performance liquid chromatography (UHPLC) with detection on a triple-quadrupole mass spectrometer operating in both positive and negative electrospray ionization modes.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Nicholson JK, Lindon JC, Holmes E (1999) ‘Metabonomics’: understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica 29:1181–1189
Nicholson JK, Lindon JC (2008) Systems biology: Metabonomics. Nature 455:1054–1056
Nicholson JK, Connelly J, Lindon JC et al (2002) Metabonomics: a platform for studying drug toxicity and gene function. Nat Rev Drug Discov 1:153–161
Theodoridis GA, Gika HG, Wilson ID (2011) Mass spectrometry-based holistic analytical approaches for metabolite profiling in systems biology studies. Mass Spectrom Rev 30:884–906
Zhou B, Xiao JF, Tuli L, Ressom HW (2012) LC-MS-based metabolomics. Mol BioSyst 8:470–481
Fiehn O, Robertson D, Griffin J et al (2007) The metabolomics standards initiative (MSI). Metabolomics 3:175–178
Theodoridis GA, Gika HG, Want EJ et al (2012) Liquid chromatography-mass spectrometry based global metabolite profiling: a review. Anal Chim Acta 711:7–16
Verpoorte R, Choi YH, Kim HK (2010) Metabolomics: will it stay? Phytochem Anal 21:2–3
Griffiths WJ, Koal T, Wang Y et al (2010) Targeted metabolomics for biomarker discovery. Angew Chem Int Ed Engl 49:5426–5445
Michopoulos F, Whalley N, Theodoridis G et al (2014) Targeted profiling of polar intracellular metabolites using ion-pair-high performance liquid chromatography and -ultra high performance liquid chromatography coupled to tandem mass spectrom. : applications to serum, urine and tissue extracts. J Chromatogr A 1349:60–68
Roberts LD, Souza AL, Gerszten RE et al Targeted metabolomics. Curr Protoc Mol Biol 98:302.1–302.24
Gika HG, Theodoridis GA, Vrhovsek U et al (2012) Quantitative profiling of polar primary metabolites using hydrophilic interaction ultrahigh performance liquid chromatography-tandem mass spectrometry. J Chromatogr A 1259:121–127
Schiesel S, Lämmerhofer M, Lindner W (2010) Multitarget quantitative metabolic profiling of hydrophilic metabolites in fermentation broths of β-lactam antibiotics production by HILIC-ESI-MS/MS. Anal Bioanal Chem 396:1655–1679
Coulier L, Bas R, Jespersen S et al (2006) Simultaneous quantitative analysis of metabolites using ion-pair liquid chromatography-electrospray ionization mass spectrometry. Anal Chem 78:6573–6582
Buescher JM, Moco S, Sauer U et al (2010) Ultrahigh performance liquid chromatography–tandem mass spectrometry method for fast and robust quantification of anionic and aromatic metabolites. Anal Chem 82:4403–4412
Virgiliou C, Sampsonidis I, Gika HG et al (2015) Development and validation of a HILIC-MS/MS multitargeted method for metabolomics applications. Electrophoresis 36:2215–2225
Sampsonidis I, Witting M, Koch W et al (2015) Computational analysis and ratiometric comparison approaches aimed to assist column selection in hydrophilic interaction liquid chromatography–tandem mass spectrometry targeted metabolomics. J Chromatogr A 1406:145–155
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Virgiliou, C., Gika, H.G., Theodoridis, G.A. (2018). HILIC-MS/MS Multi-Targeted Method for Metabolomics Applications. In: Theodoridis, G., Gika, H., Wilson, I. (eds) Metabolic Profiling. Methods in Molecular Biology, vol 1738. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7643-0_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7643-0_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7642-3
Online ISBN: 978-1-4939-7643-0
eBook Packages: Springer Protocols