Abstract
Metabolomics is the comprehensive study of metabolism, the biochemical processes that sustain life. By comparing metabolites between healthy and disease states, new insights into disease mechanisms can be uncovered. NMR is a powerful analytical method to detect and quantify metabolites. Standard one-dimensional (1D) 1H-NMR metabolite profiling is informative but challenged by significant chemical shift overlap. Multi-dimensional NMR can increase resolution, but the required long acquisition times lead to limited throughput. Non-uniform sampling (NUS) is a well-accepted mode of acquiring multi-dimensional NMR data, enabling either reduced acquisition times or increased sensitivity in equivalent time. Despite these advantages, the technique is not widely applied to metabolomics. In this study, we evaluated the utility of NUS 1H–13C heteronuclear single quantum coherence (HSQC) for semi-quantitative metabolomics. We demonstrated that NUS improved sensitivity compared to uniform sampling (US). We verified that the NUS measurement maintains linearity, making it possible to detect metabolite changes across samples and studies. Furthermore, we calculated the lower limit of detection and quantification (LOD/LOQ) of common metabolites. Finally, we demonstrate that the measurements are repeatable on the same system and across different systems. In conclusion, our results detail the analytical capability of NUS and, in doing so, empower the future use of NUS 1H–13C HSQC in metabolomic studies.
Highlights
Metabolomics is the global measurement of metabolism, the biochemical processes that allow organisms to grow, reproduce, maintain their structures, and respond to genetic and environmental factors [1,2,3]
We evaluated the utility of Non-uniform sampling (NUS) 1 H–13 C heteronuclear single quantum coherence (HSQC) for semi-quantitative metabolomics
The acquisition time required for traditional multidimensional nuclear magnetic resonance spectroscopy (NMR) represents a significant bottleneck for metabolomics
Summary
Metabolomics is the global measurement of metabolism, the biochemical processes that allow organisms to grow, reproduce, maintain their structures, and respond to genetic and environmental factors [1,2,3]. Multi-dimensional NMR spectroscopy increases resolution and reduces signal overlap by dispersing resonances into two or more chemical shift dimensions benefiting both identification and quantification. It is important to note that the metabolite concentrations in the synthetic samples were enhanced by a factor of 20 compared to authentic serum samples Both studies only focused on the identification and quantification of a few select metabolites at millimolar concentrations. The ability of NUS HSQC to accurately characterize metabolomics profiles for a chemically diverse set of metabolites over a wide concentration range was not rigorously investigated. We systematically tested the reproducibility of NUS measurements on the same magnet and across magnets to establish a set of guidelines for future NUS 1 H–13 C HSQC metabolomic studies
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