Skip to main content

Quantification of Stable Isotope Label in Metabolites via Mass Spectrometry

  • Protocol
  • First Online:
Plant Chemical Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1056))

Abstract

Isotope labelling experiments with stable or radioactive isotopes have long been an integral part of biological and medical research. Labelling experiments led to the discovery of new metabolic pathways and made it possible to calculate the fluxes responsible for a metabolic phenotype, i.e., the qualitative and quantitative composition of metabolites in a biological system. Prerequisite for efficient isotope labelling experiments is a reliable and precise method to analyze the redistribution of isotope label in a metabolic network. Here we describe the use of the CORRECTOR program, which utilizes matrix calculations to correct mass spectral data from stable isotope labelling experiments for the distorting effect of naturally occurring stable isotopes (NOIs). CORRECTOR facilitates and speeds up the routine quantification of experimentally introduced isotope label from multiple mass spectral readouts, which are generated by routine metabolite profiling when combined with stable isotope labelling experiments.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Roessner U et al (2000) Simultaneous analysis of metabolites in potato tuber by gas chromatography–mass spectrometry. Plant J 23: 131–142

    Article  PubMed  CAS  Google Scholar 

  2. Fiehn O et al (2000) Metabolite profiling for plant functional genomics. Nat Biotechnol 18:1157–1161

    Article  PubMed  CAS  Google Scholar 

  3. Lisec J et al (2006) Gas chromatography mass spectrometry-based metabolite profiling in plants. Nat Protoc 1:387–396

    Article  PubMed  CAS  Google Scholar 

  4. Erban A et al (2007) Non-supervised construction and application of mass spectral and retention time index libraries from time-of-flight GC-MS metabolite profiles. In: Weckwerth W (ed) Metabolomics: methods and protocols. Humana Press, New York, pp 19–38

    Google Scholar 

  5. Luedemann A et al (2008) TagFinder for the quantitative analysis of gas chromatography–mass spectrometry (GC-MS) based metabolite profiling experiments. Bioinformatics 24: 732–737

    Article  PubMed  CAS  Google Scholar 

  6. Lee WNP, Byerley LO, Gergner EA (1991) Mass isotopomer analysis: theoretical and practical considerations. J Mass Spectrom 30: 451–458

    Google Scholar 

  7. Fernandez CA et al (1996) Correction of 13C mass isotopomer distributions for natural stable isotope abundance. J Mass Spectrom 31:255–262

    Article  PubMed  CAS  Google Scholar 

  8. Wittmann C, Heinzle E (1999) Mass spectrometry for metabolic flux analysis. Biotechnol Bioeng 62:739–750

    Article  PubMed  CAS  Google Scholar 

  9. van Winden WA et al (2002) Correcting mass isotopomer distributions for naturally occurring isotopes. Biotechnol Bioeng 80:477–479

    Article  PubMed  Google Scholar 

  10. Wahl AS, Dauner M, Wiechert W (2003) New tools for mass isotopomer data evaluation in 13C flux analysis: mass isotope correction, data consistency checking, and precursor relationships. Biotechnol Bioeng 85:259–268

    Article  Google Scholar 

  11. Huege J et al (2011) Modulation of the major paths of carbon in photorespiratory mutants of synechocystis. PLoS One 6(1):e16278. doi:10.1371/journal.pone.0016278

    Article  PubMed  CAS  Google Scholar 

  12. Rosman KJR, Taylor PDP (1998) Isotopic compositions of the elements 1997. Pure Appl Chem 70:217–235

    Article  CAS  Google Scholar 

  13. Huege J et al (2007) GC-EI-TOF-MS analysis of in vivo carbon-partitioning into soluble metabolite pools of higher plants by monitoring isotope dilution after (13CO2)-labelling. Phytochemistry 68:2258–2272

    Article  PubMed  CAS  Google Scholar 

  14. Allen DK, Shachar-Hill Y, Ohlrogge JB (2007) Compartment-specific labelling information in metabolic flux analysis of plants. Phytochemistry 68:2197–2210

    Article  PubMed  CAS  Google Scholar 

  15. Allen DK, Ratcliffe RG (2009) Quantification of isotope label. In: Schwender J (ed) Plant metabolic networks. Springer, New York, pp 105–149

    Chapter  Google Scholar 

  16. Leimer KR, Rice RH, Gehrke CW (1977) Complete mass spectra of the per-trimethylsilylated amino acids. J Chromatogr 141:355–375

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Max Planck society and a grant from the DFG (Deutsche Forschungsgemeinschaft) to JK. The CORRECTOR software tool was supported in part by the European META-PHOR project, FOOD-CT-2006-036220, and the GoFORSYS project (http://www.goforsys.de/) funded by the Bundesministerium für Bildung und Forschung (BMBF). JH and BH were supported by BMBF grants 0315295 and 0315426A.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, New York

About this protocol

Cite this protocol

Huege, J., Goetze, J., Dethloff, F., Junker, B., Kopka, J. (2014). Quantification of Stable Isotope Label in Metabolites via Mass Spectrometry. In: Hicks, G., Robert, S. (eds) Plant Chemical Genomics. Methods in Molecular Biology, vol 1056. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-592-7_20

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-592-7_20

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-591-0

  • Online ISBN: 978-1-62703-592-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics