Advertisement

Metabolomics pp 141-155 | Cite as

Application of Liquid Chromatography-Mass Spectrometry Analysis in Metabolomics

Reversed-Phase Monolithic Capillary Chromatography and Hydrophilic Chromatography Coupled to Electrospray Ionization-Mass Spectrometry
  • Vladimir V. Tolstikov
  • Oliver Fiehn
  • Nobuo Tanaka
Part of the Methods in Molecular Biology™ book series (MIMB, volume 358)

Abstract

Analysis of the entire metabolome as the sum of all detectable components in the sample rather than analysis of each individual metabolite is performed by the metabolomics approaches. To monitor in parallel hundreds or even thousands of metabolites, high-throughput techniques are required that enable screening for relative changes rather than absolute concentrations of compounds. Most analytical techniques for profiling small molecules consist of gas chromatography (GC) or high-performance liquid chromatography (HPLC) coupled to mass spectrometry. HPLC separations are better suited for the analysis of labile and high molecular weight compounds, and for the analysis of nonvolatile polar compounds in their natural form. Although GC- and HPLC-based profiling techniques are not truly quantitative, the compounds detecting and employing the acceptable standards may compare their relative amounts. We have demonstrated that reversed-phase monolithic capillary chromatography and hydrophilic chromatography can be successfully applied for sufficient plant crude extracts separations and metabolomics studies.

Keywords

Theoretical Plate Monolithic Column Pump Flow Rate Capillary Monolithic Column Typical HILIC 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Tolstikov, V. V., Lommen, A., Nakanishi, K., Tanaka, N., and Fiehn O. (2003) Monolithic silica-based capillary reversed phase liquid chromatography/electrospray mass spectrometry for plant metabolomics. Anal. Chem. 75, 6737–6740.PubMedCrossRefGoogle Scholar
  2. 2.
    Tolstikov, V. V., Fiehn, O., and Shulaev, V. (2005) The influence of the extraction methodology on global LC-MS and GC-MS metabolite profiling of arabidopsis thaliana leaf tissues. 53rd ASMS Conference, San Antonio, TX, June 5–9, 2005.Google Scholar
  3. 3.
    Tolstikov, V. V., Lorence, A., Cortes, D. F., et al. (2005) Use of capillary monolithic columns for untargeted metabolite profiling of the mutant lines overexpressing Miox4, the gene involved in L-ascorbic acid biosynthesis in arabidopsis. 28th International Symposium On Capillary Chromatography and Electrophoresis. Las Vegas, NV, May 22–25, 2005.Google Scholar
  4. 4.
    Tolstikov, V. V. and Fiehn, O. (2002) Analysis of highly polar compounds of plant origin: combination of hydrophilic interaction chromatography and electrospray ion trap mass spectrometry. Anal. Biochem. 301, 298–307.PubMedCrossRefGoogle Scholar
  5. 5.
    Tolstikov, V. V., Tanaka, N., and Fiehn O. (2003) Comprehensive metabolome analysis of crude Arabidopsis thaliana leaf extracts by LC/ESI-MSn/UV coupling. 2nd Plant Metabolomics Conference, Potsdam, Germany, April 25–28, 2003.Google Scholar
  6. 6.
    Tolstikov, V. V., Tanaka, N., and Fiehn, O. (2003) Metabolomics: LC-MS analysis development. Joint BTS/Cereal Chemistry Symposium, Adelaide, Australia, September 8–10, 2003.Google Scholar
  7. 7.
    Tolstikov, V. V., Zhang, B., Weckwerth, W., and Fiehn O. (2001) Structural investigation of O-glycans derived from plant material by the use of the HILIC HPLC separation and ESI-mass spectrometry. 49th ASMS conference on Mass Spectrometry and Allied Topics, Chicago, IL, May 27–31, 2001.Google Scholar
  8. 8.
    Tolstikov, V. V., Costisella, B., Weckwerth W., Zhang B., and Fiehn O. (2002) Accurate QTOF and MSn Ion trap measurements require additional NMR data for plant metabolites de-novo identification. 50th ASMS Conference on Mass Spectrometry and Allied Topics, Orlando, FL, June 2–7, 2002.Google Scholar
  9. 9.
    Tolstikov, V. V., Tanaka, N., and Fiehn, O. (2003) LC/MS analysis and development for plant metabolomic studies. 2003 LCMS Montreux Symposium. Savannah, GA, October 15–17, 2003.Google Scholar
  10. 10.
    Tanaka, N., Tolstikov, V., Weckwerth, W., Fiehn, O., and Fukusaki, H., (2003) Micro HPLC for metabolomics. In: Frontier of Metabolomic Research, Springer-Verlag, Tokyo, Japan, pp. 85–100.Google Scholar
  11. 11.
    Ikegami, T., Kobayashi, H., Kimura, H., Tolstikov, V., Fiehn, O., and Tanaka, N. (2005) HPLC for metabolomics: high efficiency separations utilizing monolithic silica columns. In: Metabolomics. The Frontiers of Systems Biology, Springer Verlag, Tokyo, Japan, pp. 107–126.Google Scholar
  12. 12.
    Nikiforova, V., Kopka, J., Tolstikov, V., Fiehn, O., Hesse, H., and Hoefgen, R. (2005) Systems rebalancing of metabolism in response to sulfur deprivation, as revealed by metabolome analysis of arabidopsis plants. Plant Physiology 138, 304–318.PubMedCrossRefGoogle Scholar
  13. 13.
    Kobayashi, H., Kajiwara, W., Inui, Y., et al. (2004) Chromatographic properties of monolithic silica capillary columns for polar and nonpolar compounds in reversed-phase HPLC. Chromatographia 60, S19–S25.CrossRefGoogle Scholar
  14. 14.
    Ikegami, T., Dicks, E., Kobayashi, H., et al. (2004) How to utilize true performance of monolithic silica columns? J. Sep. Sci. 27, 1292–1302.PubMedCrossRefGoogle Scholar
  15. 15.
    Richardson, P. T. and Baker, D. A. (1982) The chemical composition of cucurbit vascular exudates. J. Exp. Bot. 33, 1239–1247.CrossRefGoogle Scholar
  16. 16.
    Snyder, L. R., Glajch, J. L., and Kirkland, J. J. (1988) Non-ionic samples: reversed-and normal-phase HPLC. In: Practical HPLC Method Development. Wiley, New York, NY, pp. 233–291.Google Scholar
  17. 17.
    Snyder, L. R., Stadalius, M., and Quarry, M. A. (1983) Gradient elution in reversed-phase HPLC separation of macromolecules. Anal. Chem. 55, 1412A–1430A.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Vladimir V. Tolstikov
    • 1
  • Oliver Fiehn
    • 1
  • Nobuo Tanaka
    • 2
  1. 1.Genome CenterUniversity of California-DavisDavis
  2. 2.Department of Polymer Science and EngineeringKyoto Institute of TechnologyKyotoJapan

Personalised recommendations