title: Annotation of the human serum metabolome by coupling three liquid chromatography methods to high-resolution mass spectrometry.
abstract: This work aims at evaluating the relevance and versatility of liquid chromatography coupled to high resolution mass spectrometry (LC/HRMS) for performing a qualitative and comprehensive study of the human serum metabolome. To this end, three different chromatographic systems based on a reversed phase (RP), hydrophilic interaction chromatography (HILIC) and a pentafluorophenylpropyl (PFPP) stationary phase were used, with detection in both positive and negative electrospray modes. LC/HRMS platforms were first assessed for their ability to detect, retain and separate 657 metabolite standards representative of the chemical families occurring in biological fluids. More than 75% were efficiently retained in either one LC-condition and less than 5% were exclusively retained by the RP column. These three LC/HRMS systems were then evaluated for their coverage of serum metabolome. The combination of RP, HILIC and PFPP based LC/HRMS methods resulted in the annotation of about 1328 features in the negative ionization mode, and 1358 in the positive ionization mode on the basis of their accurate mass and precise retention time in at least one chromatographic condition. Less than 12% of these annotations were shared by the three LC systems, which highlights their complementarity. HILIC column ensured the greatest metabolome coverage in the negative ionization mode, whereas PFPP column was the most effective in the positive ionization mode. Altogether, 192 annotations were confirmed using our spectral database and 74 others by performing MS/MS experiments. This resulted in the formal or putative identification of 266 metabolites, among which 59 are reported for the first time in human serum.
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives/Direction des Sciences du Vivant/Institut de Biologie et de Technologie de Saclay/Service de Pharmacologie et d'Immunoanalyse/Laboratoire d'Etude du Métabolisme des Médicaments, CEA-Saclay, 91191 Gif-Sur-Yvette, France; GlaxoSmithKline - Centre de recherche F.Hyafil, 25 Avenue du Québec, Villebon-sur-Yvette, France.
authors.forename: Samia
authors.initials: S
authors.lastname: Boudah
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives/Direction des Sciences du Vivant/Institut de Biologie et de Technologie de Saclay/Service de Pharmacologie et d'Immunoanalyse/Laboratoire d'Etude du Métabolisme des Médicaments, CEA-Saclay, 91191 Gif-Sur-Yvette, France.
authors.forename: Marie-Françoise
authors.initials: MF
authors.lastname: Olivier
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives/Direction des Sciences du Vivant/Institut de Biologie et de Technologie de Saclay/Service de Pharmacologie et d'Immunoanalyse/Laboratoire d'Etude du Métabolisme des Médicaments, CEA-Saclay, 91191 Gif-Sur-Yvette, France; bioMérieux S.A, Chemin de l'Orme, 69280 Marcy l'Etoile, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Aros-Calt
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives/Direction des Sciences du Vivant/Institut de Radiobiologie Cellulaire et Moléculaire/Laboratoire de Recherche sur la Transcription et la Réparation des Cellules Souches, 92260 Fontenay-aux-Roses, France.
authors.forename: Lydie
authors.initials: L
authors.lastname: Oliveira
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives/Direction des Sciences du Vivant/Institut de Biologie et de Technologie de Saclay/Service de Pharmacologie et d'Immunoanalyse/Laboratoire d'Etude du Métabolisme des Médicaments, CEA-Saclay, 91191 Gif-Sur-Yvette, France.
authors.forename: François
authors.initials: F
authors.lastname: Fenaille
authors.affiliation: Laboratoire de Chimie Structurale Organique et Biologique, IPCM/CNRS UMR 7201, Université Pierre et Marie Curie, 75252 Paris, France.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Tabet
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives/Direction des Sciences du Vivant/Institut de Biologie et de Technologie de Saclay/Service de Pharmacologie et d'Immunoanalyse/Laboratoire d'Etude du Métabolisme des Médicaments, CEA-Saclay, 91191 Gif-Sur-Yvette, France. Electronic address: christophe.junot@cea.fr.
authors.forename: Christophe
authors.initials: C
authors.lastname: Junot
mesh_terms: D001774:Blood Chemical Analysis / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D016208:Databases, Factual; D006801:Humans; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D009930:Organic Chemicals / Q000097:blood* / Q000737:chemistry / Q000145:classification; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009930:Organic Chemicals
keywords: High resolution mass spectrometry; Liquid chromatography; Metabolite identification; Metabolome annotation; Metabolomics; Serum; Tandem mass spectrometry