Abstract
Liquid chromatography-mass spectrometry (LC-MS or HPLC-MS) is an extremely sensitive analytical technique that enables the detection of metabolites with a vast range of chemistries and molecular masses. Extracts from any biological starting material are first fractionated chromatographically on a stationary phase suitable for the retention of the target molecules. The eluent is then transferred directly to the ionization source for MS detection. There is a vast range of chromatographic separation methods and MS configurations. This chapter describes a method for the detection of a broad range of metabolites using reversed phase (C18) LC-MS as well as a method for the isolation of targeted metabolites of interest.
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References
Zhou B, Xiao JF, Tuli L et al (2012) LC-MS-based metabolomics. Mol Biosyst 8:470–481
Watson JT, Sparkman OD (2008) Introduction to mass spectrometry. Instrumentation, applications and strategies for data interpretation, 4th edn. Wiley, Chichester, NY
Nakabayashi R, Kusano M, Kobayashi M et al (2009) Metabolomics-oriented isolation and structure elucidation of 37 compounds including two anthocyanins from Arabidopsis thaliana. Phytochemistry 70:1017–1029
Vuckovic D (2012) Current trends and challenges in sample preparation for global metabolomics using liquid chromatography-mass spectrometry. Anal Bioanal Chem 403:1523–1548
Lu W, Bennett BD, Rabinowitz JD (2009) Analytical strategies for LC-MS-based targeted metabolomics. J Chromatogr B Analyt Technol Biomed Life Sci 871:236–242
Ellis DI, Dunn WB, Griffin JL et al (2007) Metabolic fingerprinting as a diagnostic tool. Pharmacogenomics 8:1243–1266
De Vos RCH, Moco S, Lommen A et al (2007) Untargeted large-scale plant metabolomics using liquid chromatography coupled to mass spectrometry. Nat Protoc 2:778–791
Rojo D, Barbas C, Ruperez FJ (2012) LC-MS metabolomics of polar compounds. Bioanalysis 4:1235–1243
Keller BO, Sui J, Young AB et al (2008) Interferences and contaminants encountered in modern mass spectrometry. Anal Chim Acta 627:71–81
Zelena E, Dunn WB, Broadhurst D et al (2009) Development of a robust and repeatable UPLC − MS method for the long-term metabolomic study of human serum. Anal Chem 81:1357–1364
van den Berg R, Hoefsloot H, Westerhuis J et al (2006) Centering, scaling, and transformations: improving the biological information content of metabolomics data. BMC Genomics 7:142
Pluskal T, Castillo S, Villar-Briones A et al (2010) MZmine 2: modular framework for processing, visualizing, and analyzing mass spectrometry-based molecular profile data. BMC Bioinformatics 11:1471–2105
Smith CA, Want EJ, O’Maille G et al (2006) XCMS: processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification. Anal Chem 78:779–787
Lommen A, Kools H (2012) MetAlign 3.0: performance enhancement by efficient use of advances in computer hardware. Metabolomics 8:719–726
Howlett BJ, Idnurm A, Pedras MS (2001) Leptosphaeria maculans, the causal agent of blackleg disease of Brassicas. Fungal Genet Biol 33:1–14
Fiehn O, Sumner L, Ward J, Rhee SY, Dickerson J, Lange M, Lane G, Roessner U, Last R, Nikolau B (2007) Minimum reporting standards for plan biology context in metabolomic studies. Metabolomics 3, 195–201
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Callahan, D.L., Elliott, C.E. (2013). A Workflow from Untargeted LC-MS Profiling to Targeted Natural Product Isolation. In: Roessner, U., Dias, D. (eds) Metabolomics Tools for Natural Product Discovery. Methods in Molecular Biology, vol 1055. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-577-4_5
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DOI: https://doi.org/10.1007/978-1-62703-577-4_5
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