Rapid two-dimensional ALSOFAST-HSQC experiment for metabolomics and fluxomics studies: application to a13 C-enriched cancer cell model treated with gold nanoparticles
Abstract
Isotope labeling enables the use of13C-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-dimensional13C and classical two-dimensional1H,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 of13C-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 on13C 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 various13C-enriched metabolites and the determination of specific variations between the different sample groups. Thus, it was possible...Continue Reading
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