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Rapid two-dimensional ALSOFAST-HSQC experiment for metabolomics and fluxomics studies: application to a 13C-enriched cancer cell model treated with gold nanoparticles

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Abstract

Isotope labeling enables the use of 13C-based metabolomics techniques with strongly improved resolution for a better identification of relevant metabolites and tracing of metabolic fluxes in cell and animal models, as required in fluxomics studies. However, even at high NMR-active isotope abundance, the acquisition of one-dimensional 13C and classical two-dimensional 1H,13C-HSQC experiments remains time consuming. With the aim to provide a shorter, more efficient alternative, herein we explored the ALSOFAST-HSQC experiment with its rapid acquisition scheme for the analysis of 13C-labeled metabolites in complex biological mixtures. As an initial step, the parameters of the pulse sequence were optimized to take into account the specific characteristics of the complex samples. We then applied the fast two-dimensional experiment to study the effect of different kinds of antioxidant gold nanoparticles on a HeLa cancer cell model grown on 13C glucose-enriched medium. As a result, 1H,13C-2D correlations could be obtained in a couple of seconds to few minutes, allowing a simple and reliable identification of various 13C-enriched metabolites and the determination of specific variations between the different sample groups. Thus, it was possible to monitor glucose metabolism in the cell model and study the antioxidant effect of the coated gold nanoparticles in detail. Finally, with an experiment time of only half an hour, highly resolved 1H,13C-HSQC spectra using the ALSOFAST-HSQC pulse sequence were acquired, revealing the isotope-position-patterns of the corresponding 13C-nuclei from carbon multiplets.

Fast NMR applied to metabolomics and fluxomics studies with gold nanoparticles

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Acknowledgements

Part of the work was supported by grants CP13/00252 and PI16/02064 from Carlos III Health Institute, SAF2014-53977-R and SAF2017-89229-R from Ministerio de Economía y Competitividad, and by the European Regional Development Fund (ERDF). M.P.-S. was partially supported by an EMBO short-term fellowship. Eva Castelló has contributed to the work with technical help. B.L. thanks the HGF programme BIFTM, the DFG (instrumentation facility Pro2NMR and LU 835/13-1), and Fonds der Chemischen Industrie for financial support. We also want to thank Martin Koos for helpful discussions.

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Authors and Affiliations

  1. Institut für Organische Chemie, Karlsruher Institut für Technologie (KIT), Fritz-Haber-Weg 6, 76131, Karlsruhe, Germany

    Martina Palomino Schätzlein, Johanna Becker, David Schulze-Sünninghausen & Burkhard Luy

  2. Institut für Biologische Grenzflächen 4 - Magnetische Resonanz, Karlsruher Institut für Technologie (KIT), Postfach 3640, 76021, Karlsruhe, Germany

    Martina Palomino Schätzlein

  3. Centro de Investigación Príncipe Felipe, Calle Eduardo Primo Yúfera 3, 46012, Valencia, Spain

    Martina Palomino Schätzlein & Antonio Pineda-Lucena

  4. Bruker BioSpin GmbH, Rudolf-Planck-Str. 23, 76275, Ettlingen, Germany

    David Schulze-Sünninghausen

  5. Drug Discovery Unit, Instituto de Investigación Sanitaria La Fe, Avda. Fernando Abril Martorell, 106, 46026, Valencia, Spain

    Antonio Pineda-Lucena

  6. Medical Molecular Imaging Research Group, Vall d’Hebron Research Institute, CIBBIM-Nanomedicine, Pg. Vall d’Hebron 119-129, 08035, Barcelona, Spain

    José Raul Herance

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  1. Martina Palomino Schätzlein
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Correspondence to Martina Palomino Schätzlein or Burkhard Luy.

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Schätzlein, M.P., Becker, J., Schulze-Sünninghausen, D. et al. Rapid two-dimensional ALSOFAST-HSQC experiment for metabolomics and fluxomics studies: application to a 13C-enriched cancer cell model treated with gold nanoparticles. Anal Bioanal Chem 410, 2793–2804 (2018). https://doi.org/10.1007/s00216-018-0961-6

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