title: A multiplexed targeted assay for high-throughput quantitative analysis of serum methylamines by ultra performance liquid chromatography coupled to high resolution mass spectrometry.
abstract: Methylamines are biologically-active metabolites present in serum and urine samples, which play complex roles in metabolic diseases. Methylamines can be detected by proton nuclear magnetic resonance (NMR), but specific methods remain to be developed for their routine assay in human serum in clinical settings. Here we developed and validated a novel reliable "methylamine panel" method for simultaneous quantitative analysis of trimethylamine (TMA), its major detoxification metabolite trimethylamine-N-oxide (TMAO), and precursors choline, betaine and l-carnitine in human serum using Ultra Performance Liquid Chromatography (UPLC) coupled to High Resolution Mass Spectrometry (HRMS). Metabolite separation was carried out on a HILIC stationary phase. For all metabolites, the assay was linear in the range of 0.25-12.5 μmol/L and enabled to reach limit of detection of about 0.10 μmol/L. Relative standard deviations were below 16% for the three levels of concentrations. We demonstrated the strong reliability and robustness of the method, which was applied to serum samples from healthy individuals to establish the range of concentrations of the metabolites and their correlation relationships and detect gender differences. Our data provide original information for implementing in a clinical environment a MS-based diagnostic method with potential for targeted metabolic screening of patients at risk of cardiometabolic diseases.
authors.affiliation: Sorbonne Universities, University Pierre & Marie Curie, University Paris Descartes, Sorbonne Paris Cité, INSERM, UMR_S 1138, Cordeliers Research Centre, 15 rue de l'Ecole de Médecine, 75006, Paris, France; Institute of Cardiometabolism and Nutrition, University Pierre & Marie Curie, Hospital Pitié Salpetrière, 58 Boulevard de l'hôpital, 75013, Paris, France.
authors.forename: Hanane
authors.initials: H
authors.lastname: Kadar
authors.affiliation: Institute of Cardiometabolism and Nutrition, University Pierre & Marie Curie, Hospital Pitié Salpetrière, 58 Boulevard de l'hôpital, 75013, Paris, France.
authors.forename: Justine
authors.initials: J
authors.lastname: Dubus
authors.affiliation: Sorbonne Universities, University Pierre & Marie Curie, University Paris Descartes, Sorbonne Paris Cité, INSERM, UMR_S 1138, Cordeliers Research Centre, 15 rue de l'Ecole de Médecine, 75006, Paris, France.
authors.forename: Jérémie
authors.initials: J
authors.lastname: Dutot
authors.affiliation: Institute of Cardiometabolism and Nutrition, University Pierre & Marie Curie, Hospital Pitié Salpetrière, 58 Boulevard de l'hôpital, 75013, Paris, France.
authors.forename: Lyamine
authors.initials: L
authors.lastname: Hedjazi
authors.affiliation: Program in Nutritional Metabolism, Massachusetts General Hospital and Harvard Medical School, Boston, MA, 02114, United States.
authors.forename: Suman
authors.initials: S
authors.lastname: Srinivasa
authors.affiliation: Program in Nutritional Metabolism, Massachusetts General Hospital and Harvard Medical School, Boston, MA, 02114, United States.
authors.forename: Kathleen V
authors.initials: KV
authors.lastname: Fitch
authors.affiliation: Program in Nutritional Metabolism, Massachusetts General Hospital and Harvard Medical School, Boston, MA, 02114, United States.
authors.forename: Steven K
authors.initials: SK
authors.lastname: Grinspoon
authors.affiliation: Imperial College London, Section of Biomolecular Medicine, Division of Computational Systems Medicine, Department of Surgery and Cancer, Sir Alexander Fleming Building, Exhibition Road, South Kensington, London, SW7 2AZ, United Kingdom; Imperial College London, MRC-NIHR National Phenome Centre, Department of Surgery and Cancer, IRDB Building, Du Cane Road, London, W12 0NN, United Kingdom.
authors.forename: Jeremy K
authors.initials: JK
authors.lastname: Nicholson
authors.affiliation: Imperial College London, Section of Biomolecular Medicine, Division of Computational Systems Medicine, Department of Surgery and Cancer, Sir Alexander Fleming Building, Exhibition Road, South Kensington, London, SW7 2AZ, United Kingdom; Imperial College London, MRC-NIHR National Phenome Centre, Department of Surgery and Cancer, IRDB Building, Du Cane Road, London, W12 0NN, United Kingdom. Electronic address: m.dumas@imperial.ac.uk.
authors.forename: Marc-Emmanuel
authors.initials: ME
authors.lastname: Dumas
authors.affiliation: Sorbonne Universities, University Pierre & Marie Curie, University Paris Descartes, Sorbonne Paris Cité, INSERM, UMR_S 1138, Cordeliers Research Centre, 15 rue de l'Ecole de Médecine, 75006, Paris, France; Imperial College London, Section of Biomolecular Medicine, Division of Computational Systems Medicine, Department of Surgery and Cancer, Sir Alexander Fleming Building, Exhibition Road, South Kensington, London, SW7 2AZ, United Kingdom. Electronic address: dominique.gauguier@crc.jussieu.fr.
authors.forename: Dominique
authors.initials: D
authors.lastname: Gauguier
mesh_terms: D000328:Adult; D015415:Biomarkers; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008744:Methylamines / Q000097:blood*; D008875:Middle Aged
chemical_list: D015415:Biomarkers; D008744:Methylamines
keywords: Betaine; Cardiometabolic diseases; Choline; Trimethylamine; Trimethylamine-N-oxide; l-carnitine