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Quantitative Metabolomic Profiling Using Dansylation Isotope Labeling and Liquid Chromatography Mass Spectrometry

  • Ruokun Zhou
  • Liang Li
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1198)

Abstract

Differential chemical isotopic labeling (CIL) LC-MS has been used for quantifying a targeted metabolite in biological samples with high precision and accuracy. Herein we describe a high-performance CIL LC-MS method for generating quantitative and comprehensive profiles of the metabolome for metabolomics applications. After mixing two comparative samples separately labeled by light or heavy isotopic tags through chemical reactions, the peak intensity ratio of the labeled analyte pair can provide relative or absolute quantitative information on the metabolites. We describe the use of 12C2- and 13C2-dansyl chloride (DnsCl) as the isotope reagents to profile the metabolites containing amine and phenolic hydroxyl functional groups by LC-MS. This method can be used to compare the relative concentration changes of hundreds or thousands of amine- and phenol-containing metabolites among many comparative samples and generate absolute concentration information on metabolites for which the standards are available. Combined with statistical analysis and metabolite identification tools, this method can be used to identify key metabolites involved in differentiating comparative samples such as disease cases vs. healthy controls.

Key words

Isotopic labeling reagent Dansyl chloride (DnsCl) Dansylation LC-MS Metabolomic profiling Data analysis 

Notes

Acknowledgements

This work was supported by Genome Canada, the Natural Sciences and Engineering Research Council of Canada (NSERC), Canadian Institutes of Health Research, Alberta Innovates, and Canada Research Chairs programs (CRC).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of ChemistryUniversity of AlbertaEdmontonCanada

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