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A comparison of high-throughput plasma NMR protocols for comparative untargeted metabolomics.

ABSTRACT: INTRODUCTION:When analyzing the human plasma metabolome with Nuclear Magnetic Resonance (NMR) spectroscopy, the Carr-Purcell-Meiboom-Gill (CPMG) experiment is commonly employed for large studies. However, this process can lead to compromised statistical analyses due to residual macromolecule signals. In addition, the utilization of Trimethylsilylpropanoic acid (TSP) as an internal standard often leads to quantification issues, and binning, as a spectral summarization step, can result in features not clearly assignable to metabolites. OBJECTIVES:Our aim was to establish a new complete protocol for large plasma cohorts collected with the purpose of describing the comparative metabolic profile of groups of samples. METHODS:We compared the conventional CPMG approach to a novel procedure that involves diffusion NMR, using the Longitudinal Eddy-Current Delay (LED) experiment, maleic acid (MA) as the quantification reference and peak picking for spectral reduction. This comparison was carried out using the ultrafiltration method as a gold standard in a simple sample classification experiment, with Partial Least Squares-Discriminant Analysis (PLS-DA) and the resulting metabolic signatures for multivariate data analysis. In addition, the quantification capabilities of the method were evaluated. RESULTS:We found that the LED method applied was able to detect more metabolites than CPMG and suppress macromolecule signals more efficiently. The complete protocol was able to yield PLS-DA models with enhanced classification accuracy as well as a more reliable set of important features than the conventional CPMG approach. Assessment of the quantitative capabilities of the method resulted in good linearity, recovery and agreement with an established amino acid assay for the majority of the metabolites tested. Regarding repeatability,?~?85% of all peaks had an adequately low coefficient of variation (

SUBMITTER: Bliziotis NG

PROVIDER: S-EPMC7196944 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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A comparison of high-throughput plasma NMR protocols for comparative untargeted metabolomics.

Bliziotis Nikolaos G NG Engelke Udo F H UFH Aspers Ruud L E G RLEG Engel Jasper J Deinum Jaap J Timmers Henri J L M HJLM Wevers Ron A RA Kluijtmans Leo A J LAJ

Metabolomics : Official journal of the Metabolomic Society 20200501 5

<h4>Introduction</h4>When analyzing the human plasma metabolome with Nuclear Magnetic Resonance (NMR) spectroscopy, the Carr-Purcell-Meiboom-Gill (CPMG) experiment is commonly employed for large studies. However, this process can lead to compromised statistical analyses due to residual macromolecule signals. In addition, the utilization of Trimethylsilylpropanoic acid (TSP) as an internal standard often leads to quantification issues, and binning, as a spectral summarization step, can result in ...[more]

PMID: 32358672

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A comparison of high-throughput plasma NMR protocols for comparative untargeted metabolomics.

Publications

A comparison of high-throughput plasma NMR protocols for comparative untargeted metabolomics.

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