NMR-Based Metabolic Profiling Procedures for Biofluids and Cell and Tissue Extracts

  • Dimitra Benaki
  • Emmanuel Mikros
Part of the Methods in Molecular Biology book series (MIMB, volume 1738)


Metabolomic studies offer a wealth of information on cells, tissues, and biofluids. The phenotype representation through the metabolic profiling is a valuable tool for direct diagnosis, therapeutic strategies, and system’s biology studies. Nuclear magnetic resonance (NMR) spectroscopy provides a nondestructive and extremely reproducible method allowing simultaneous detection of a large number of known and unknown chemical substances.

Sample collection and preparation and experimental conditions are critical for the reliability of the subsequent analysis. The pre-analytical phase is decisive as it could generate biased spectral data misleading the following analysis. The formulation of standard operating procedures is thus of crucial importance in order to access meaningful samples and results. In this protocol, we provide standardized operations and routine procedures from sample preparation to determine the measurement details for the acquisition of NMR spectra highlighting major methodological issues.

Key words

NMR spectroscopy Metabolic profiling Biofluids Cell cultures Tissues 



The authors wish to acknowledge Eberhard Humpfer and Manfred Spraul (Bruker BioSpin, Karlsruhe) for useful help and advice on NMR parameters optimization.


  1. 1.
    Chen R, Mias GI, Li-Pook-Than J et al (2012) Personal omics profiling reveals dynamic molecular and medical phenotypes. Cell 148(6):1293–1307. Cohen J (March, 2012). Examining his own body, stanford geneticist stops diabetes in its tracks. Retrieved from CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Dietmair S, Timmins NE, Gray PP et al (2010) Towards quantitative metabolomics of mammalian cells: development of a metabolite extraction protocol. Anal Biochem 404:155–164. CrossRefPubMedGoogle Scholar
  3. 3.
    Collinet H, Renault D (2012) Metabolic effects of CO2 anaesthesia in Drosophila Melanogaster. Biol Lett 8:1050–1054. CrossRefGoogle Scholar
  4. 4.
    Ghini V, Unger FT, Tenori L et al (2015) Metabolomics profiling of pre-and post-anesthesia plasma samples of colorectal patients obtained via Ficoll separation. Metabolomics 11:1769–1778. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Overmyer KA, Thonusin C, Qi NR et al (2015) Impact of anesthesia and euthanasia on metabolomics of mammalian tissues: studies in a C57BL/6J mouse model. PLoS One 10(2):e0117232. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Nicholson JK, Buckingham MJ, Sadler PJ (1983) High resolution 1H NMR studies of vertebrate blood and plasma. Biochem J 211(3):605–615CrossRefGoogle Scholar
  7. 7.
    Keun HC, Athersuch TJ (2011) Nuclear magnetic resonance (NMR)-based metabolomics. In: Metz TO (ed) Metabolic profiling, Methods in molecular biology, vol vol 708. Springer Protocols, Humana Press, New York, pp 321–334. CrossRefGoogle Scholar
  8. 8.
    Beckonert O, Keun HC, Ebels TMD et al (2007) Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts. Nat Protoc 2(11):2692–2703. CrossRefPubMedGoogle Scholar
  9. 9.
    Sapcariu SC, Kanashova T, Weindl D et al (2014) Simultaneous extraction of proteins and metabolites from cells in culture. MethodsX 1:74–80. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Le Belle JE, Harris NG, Williams SR et al (2002) A comparison of cell and tissue extraction techniques using high-resolution 1H-NMR spectroscopy. NMR Biomed 15:37–44CrossRefGoogle Scholar
  11. 11.
    Gowda NGA, Raftery D (2014) Quantitating metabolites in protein precipitated serum using NMR spectroscopy. Anal Chem 86(11):5433–5440. CrossRefPubMedCentralGoogle Scholar
  12. 12.
    Kapoore RV, Coyle R, Staton CA et al (2015) Cell line dependence of metabolite leakage in metabolome analyses of adherent normal and cancer cell lines. Metabolomics 11:1743–1755. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmaceutical ChemistryNational and Kapodistrian University of AthensAthensGreece

Personalised recommendations