title: Optimization and Application of Direct Infusion Nanoelectrospray HRMS Method for Large-Scale Urinary Metabolic Phenotyping in Molecular Epidemiology.
abstract: Large-scale metabolic profiling requires the development of novel economical high-throughput analytical methods to facilitate characterization of systemic metabolic variation in population phenotypes. We report a fit-for-purpose direct infusion nanoelectrospray high-resolution mass spectrometry (DI-nESI-HRMS) method with time-of-flight detection for rapid targeted parallel analysis of over 40 urinary metabolites. The newly developed 2 min infusion method requires <10 μL of urine sample and generates high-resolution MS profiles in both positive and negative polarities, enabling further data mining and relative quantification of hundreds of metabolites. Here we present optimization of the DI-nESI-HRMS method in a detailed step-by-step guide and provide a workflow with rigorous quality assessment for large-scale studies. We demonstrate for the first time the application of the method for urinary metabolic profiling in human epidemiological investigations. Implementation of the presented DI-nESI-HRMS method enabled cost-efficient analysis of >10 000 24 h urine samples from the INTERMAP study in 12 weeks and >2200 spot urine samples from the ARIC study in <3 weeks with the required sensitivity and accuracy. We illustrate the application of the technique by characterizing the differences in metabolic phenotypes of the USA and Japanese population from the INTERMAP study.
authors.affiliation: Biomolecular Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London , South Kensington, London SW7 2AZ, United Kingdom.
authors.forename: Elena
authors.identifier: 0000-0003-1807-2398
authors.initials: E
authors.lastname: Chekmeneva
authors.affiliation: Biomolecular Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London , South Kensington, London SW7 2AZ, United Kingdom.
authors.forename: Gonçalo
authors.identifier: 0000-0001-8271-9294
authors.initials: G
authors.lastname: Dos Santos Correia
authors.affiliation: Department of Epidemiology and Biostatistics, School of Public Health, Faculty of Medicine, Imperial College London , London W2 1PG, United Kingdom.|MRC-PHE Centre for Environment and Health, Imperial College London , London W2 1PG, United Kingdom.
authors.forename: Queenie
authors.initials: Q
authors.lastname: Chan
authors.affiliation: Biomolecular Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London , South Kensington, London SW7 2AZ, United Kingdom.
authors.forename: Anisha
authors.identifier: 0000-0001-6151-5065
authors.initials: A
authors.lastname: Wijeyesekera
authors.affiliation: Department of Medicine, Johns Hopkins University, School of Medicine , Baltimore, Maryland 21287, United States.
authors.forename: Adrienne
authors.initials: A
authors.lastname: Tin
authors.affiliation: Department of Medicine, Johns Hopkins University, School of Medicine , Baltimore, Maryland 21287, United States.
authors.forename: Jeffery Hunter
authors.initials: JH
authors.lastname: Young
authors.affiliation: Department of Epidemiology and Biostatistics, School of Public Health, Faculty of Medicine, Imperial College London , London W2 1PG, United Kingdom.|MRC-PHE Centre for Environment and Health, Imperial College London , London W2 1PG, United Kingdom.|MRC-NIHR National Phenome Centre , London SW7 2AZ, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Elliott
authors.affiliation: Biomolecular Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London , South Kensington, London SW7 2AZ, United Kingdom.|MRC-NIHR National Phenome Centre , London SW7 2AZ, United Kingdom.
authors.forename: Jeremy K
authors.initials: JK
authors.lastname: Nicholson
authors.affiliation: Biomolecular Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London , South Kensington, London SW7 2AZ, United Kingdom.|MRC-NIHR National Phenome Centre , London SW7 2AZ, United Kingdom.
authors.forename: Elaine
authors.identifier: 0000-0002-0556-8389
authors.initials: E
authors.lastname: Holmes
mesh_terms: D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D055442:Metabolome / Q000235:genetics*; D055432:Metabolomics / Q000379:methods; D017720:Molecular Epidemiology / Q000379:methods*; D036103:Nanotechnology / Q000379:methods; D014556:Urine / Q000737:chemistry*
keywords: direct infusion mass spectrometry; high-throughput analysis; metabolic profiling; molecular epidemiology