In this community effort, we compare measurements between 34 laboratories from 19 countries, utilizing mixtures of labelled authentic synthetic standards, to quantify by mass spectrometry four clinically used ceramide species in the NIST (National Institute of Standards and Technology) human blood plasma Standard Reference Material (SRM) 1950, as well as a set of candidate plasma reference materials (RM 8231). Participants either utilized a provided validated method and/or their method of choice. Mean concentration values, and intra- and inter-laboratory coefficients of variation (CV) were calculated using single-point and multi-point calibrations, respectively. These results are the most precise (intra-laboratory CVs ≤ 4.2%) and concordant (inter-laboratory CVs < 14%) community-derived absolute concentration values reported to date for four clinically used ceramides in the commonly analyzed SRM 1950. We demonstrate that calibration using authentic labelled standards dramatically reduces data variability. Furthermore, we show how the use of shared RM can correct systematic quantitative biases and help in harmonizing lipidomics. Collectively, the results from the present study provide a significant knowledge base for translation of lipidomic technologies to future clinical applications that might require the determination of reference intervals (RIs) in various human populations or might need to estimate reference change values (RCV), when analytical variability is a key factor for recall during multiple testing of individuals.

ArcticMass Reykjavik Iceland

Avanti Polar Lipids Alabaster AL USA

Babraham Institute Babraham Research Campus Cambridge MA CB22 3AT USA

Baker Department of Cardiovascular Research Translation and Implementation La Trobe University Bundoora VIC 3086 Australia

Baker Heart and Diabetes Institute Melbourne VIC 3004 Australia

Center for Biotechnology and Biomedicine University of Leipzig 04013 Leipzig Germany

Center for Proteomics and Metabolomics Leiden University Medical Center 2333ZA Leiden The Netherlands

Center of Membrane Biochemistry and Lipid Research Faculty of Medicine Carl Gustav Carus of TU Dresden 01307 Dresden Germany

Centro de Investigaciones en Bionanociencias Godoy Cruz 2390 C1425FQD Ciudad de Buenos Aires Argentina

Chemical Science Division National Institute of Standards and Technology Charleston SC 29412 USA

Chemical Science Division National Institute of Standards and Technology Gaithersburg MD 20899 USA

Christian Doppler Laboratory for Innovative Gut Health Concepts of Livestock Institute of Bioanalytics and Agro Metabolomics Department of Agrobiotechnology Konrad Lorenz Str 20 3430 Tulln Austria

College of Health and Life Sciences Hamad Bin Khalifa University Doha Qatar

Consejo Nacional de Investigaciones Científicas y Técnicas Departamento de Desarrollo Analítico y Control de Procesos Instituto Nacional de Tecnología Industrial Av General Paz 5445 B1650WAB Buenos Aires Argentina

Core Facility Mass Spectrometry Medical University of Graz 8010 Graz Austria

Departamento de Fisiología Biología Molecular y Celular Facultad de Ciencias Exactas y Naturales Universidad de Buenos Aires Intendente Güiraldes 2160 C1428EGA Buenos Aires Argentina

Department of Analytical Chemistry Faculty of Chemical Technology University of Pardubice Studentská 573 532 10 Pardubice Czech Republic

Department of Analytical Chemistry University of Vienna Vienna Austria

Department of Biotechnology and Life Science Tokyo University of Agriculture and Technology Tokyo Japan

Department of Chemistry University of Turku Turku Finland

Department of Chemistry Yonsei University Seoul 03722 South Korea

Department of Integrative Genomics Tohoku Medical Megabank Organization Tohoku University 2 1 Seiryo machi Aoba ku Sendai Miyagi 980 8573 Japan

Department of Pathology and Laboratory Medicine University of British Columbia Vancouver BC Canada

Department of Physiological Sciences College of Veterinary Medicine University of Florida Gainesville FL 32610 USA

Department of Respiratory Medicine and Allergy Karolinska University Hospital Stockholm Sweden

Division of Metabolomics Center Medical Research Center for High Depth Omics Medical Institute of Bioregulation Kyushu University 3‑1‑1 Maidashi Higashi‑ku Fukuoka 812‑8582 Japan

Faculty of Pharmaceutical Sciences University of Iceland Reykjavik Iceland

Graduate School of Medical Life Science Yokohama City University Yokohama Japan

Heidelberg University Biochemistry Center Im Neuenheimer Feld 328 69120 Heidelberg Germany

Institute for Bio and Geosciences Forschungszentrum Jülich GmbH 52428 Jülich Germany

Institute for Stem Cell Science and Regenerative Medicine 560065 Bangalore India

Institute of Bioanalytical Chemistry Faculty of Chemistry and Mineralogy University of Leipzig 04013 Leipzig Germany

Institute of Clinical Chemistry University Zurich 8952 Schlieren Switzerland

Laboratory for Lipidomics and Lipid Biology Division of Pharmacy and Optometry School of Health Sciences Faculty of Biology Medicine and Health The University of Manchester Manchester Academic Health Science Centre Manchester M13 9NT United Kingdom

Laboratory of Biomedical and Analytical Sciences Faculty of Pharma Science Teikyo University 2 11 1 Kaga Itabashi ku Tokyo 173 8605 Japan

Lee Kong Chian School of Medicine Nanyang Technological University Singapore 636921 Singapore

LipidALL Technologies Changzhou 213000 Jiangshu China

Lipidomics Consulting Ltd Espoo Finland

Lydia Becker Institute of Immunology and Inflammation; Faculty of Biology Medicine and Health The University of Manchester Manchester Academic Health Science Centre Manchester M13 9NT United Kingdom

MetaboHUBMetaToul Facility I2MC U1297 Inserm Toulouse France

Metabolomics and Analytics Centre Leiden Academic Centre for Drug Research Leiden University Leiden The Netherlands

Metabolomics and Proteomics Core Helmholtz Zentrum München German Research Center for Environmental Health Neuherberg 85764 Germany

Metabolomics Core Facility MetCore Universidad de los Andes Bogotá 111711 Colombia

Metabolomics Platform Faculty of Biology and Medicine University of Lausanne Lausanne Switzerland

National Centre for Biological Sciences Tata Institute of Fundamental Research Bangalore Karnataka 560065 India

Neural Regeneration Laboratory Ottawa Institute of Systems Biology Ottawa Brain and Mind Research Institute Department of Biochemistry Microbiology and Immunology and Department of Chemistry Centre for Catalysis Research and Innovation University of Ottawa Ottawa K1H 8M5 Canada

Precision Medicine Translational Research Programme and Department of Biochemistry Yong Loo Lin School of Medicine National University of Singapore Singapore 119077 Singapore

RIKEN Center for Integrative Medical Sciences Yokohama Japan

RIKEN Center for Sustainable Resource Science Yokohama Japan

School of Medical Sciences Faculty of Medicine and Health Örebro University 702 81 Örebro Sweden

Signature Research Program in Cardiovascular and Metabolic Disorders Duke National University of Singapore Medical School Singapore 169857 Singapore

Singapore Lipidomics Incubator Life Sciences Institute National University of Singapore Singapore 117456 Singapore

St Paul's Hospital Department of Pathology and Laboratory Medicine Vancouver BC Canada

State Key Laboratory of Medical Proteomics CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian 116023 China

State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China

Systems Immunity Research Institute School of Medicine Cardiff University Heath Park Cardiff CF14 4XN UK

Turku Bioscience Centre University of Turku and Åbo Akademi University 20520 Turku Finland

Unit of Integrative Metabolomics Institute of Environmental Medicine Karolinska Institutet Stockholm Sweden

Universidad de Buenos Aires Facultad de Farmacia y Bioquímica Departamento de Ciencias Químicas Buenos Aires Junin 954 Junin C1113AAD CABA Argentina

Université Paris Saclay CEA INRAE Département Médicaments et Technologies pour la Santé MetaboHUB F 91191 Gif sur Yvette France

University Hospital of Regensburg Institute of Clinical Chemistry and Laboratory Medicine Franz Josef Strauß Allee 11 93053 Regensburg Germany

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