title: High-resolution quantitative metabolome analysis of urine by automated flow injection NMR.
abstract: Metabolism is essential to understand human health. To characterize human metabolism, a high-resolution read-out of the metabolic status under various physiological conditions, either in health or disease, is needed. Metabolomics offers an unprecedented approach for generating system-specific biochemical definitions of a human phenotype through the capture of a variety of metabolites in a single measurement. The emergence of large cohorts in clinical studies increases the demand of technologies able to analyze a large number of measurements, in an automated fashion, in the most robust way. NMR is an established metabolomics tool for obtaining metabolic phenotypes. Here, we describe the analysis of NMR-based urinary profiles for metabolic studies, challenged to a large human study (3007 samples). This method includes the acquisition of nuclear Overhauser effect spectroscopy one-dimensional and J-resolved two-dimensional (J-Res-2D) (1)H NMR spectra obtained on a 600 MHz spectrometer, equipped with a 120 μL flow probe, coupled to a flow-injection analysis system, in full automation under the control of a sampler manager. Samples were acquired at a throughput of ~20 (or 40 when J-Res-2D is included) min/sample. The associated technical analysis error over the full series of analysis is 12%, which demonstrates the robustness of the method. With the aim to describe an overall metabolomics workflow, the quantification of 36 metabolites, mainly related to central carbon metabolism and gut microbial host cometabolism, was obtained, as well as multivariate data analysis of the full spectral profiles. The metabolic read-outs generated using our analytical workflow can therefore be considered for further pathway modeling and/or biological interpretation.
authors.affiliation: BioAnalytical Science, Nestle Research Center, Vers-chez-les-Blanc, P.O. Box 44, 1000 Lausanne 26, Switzerland.
authors.forename: Laeticia
authors.initials: L
authors.lastname: Da Silva
authors.forename: Markus
authors.initials: M
authors.lastname: Godejohann
authors.forename: François-Pierre J
authors.initials: FP
authors.lastname: Martin
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Collino
authors.forename: Alexander
authors.initials: A
authors.lastname: Bürkle
authors.forename: María
authors.initials: M
authors.lastname: Moreno-Villanueva
authors.forename: Jürgen
authors.initials: J
authors.lastname: Bernhardt
authors.forename: Olivier
authors.initials: O
authors.lastname: Toussaint
authors.forename: Beatrix
authors.initials: B
authors.lastname: Grubeck-Loebenstein
authors.forename: Efstathios S
authors.initials: ES
authors.lastname: Gonos
authors.forename: Ewa
authors.initials: E
authors.lastname: Sikora
authors.forename: Tilman
authors.initials: T
authors.lastname: Grune
authors.forename: Nicolle
authors.initials: N
authors.lastname: Breusing
authors.forename: Claudio
authors.initials: C
authors.lastname: Franceschi
authors.forename: Antti
authors.initials: A
authors.lastname: Hervonen
authors.forename: Manfred
authors.initials: M
authors.lastname: Spraul
authors.forename: Sofia
authors.initials: S
authors.lastname: Moco
mesh_terms: D000328:Adult; D000368:Aged; D057205:Automation, Laboratory / Q000379:methods* / Q000592:standards; D005260:Female; D017022:Flow Injection Analysis / Q000379:methods / Q000592:standards; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods* / Q000592:standards; D008297:Male; D055442:Metabolome / Q000502:physiology*; D008875:Middle Aged; D016482:Urinalysis / Q000379:methods* / Q000592:standards