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Abstract
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.
Here, the authors compared measurements between 34 laboratories from 19 countries, to quantify by mass spectrometry four ceramides of clinical relevance in human blood plasma Standard Reference Materials. The main goals were to evaluate concordance obtained in a large inter-laboratory trial and to report absolute concentrations of four circulating lipids in a publicly available standard.
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1 National University of Singapore, Singapore Lipidomics Incubator, Life Sciences Institute, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431); National University of Singapore, Precision Medicine Translational Research Programme and Department of Biochemistry, Yong Loo Lin School of Medicine, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431); Duke-National University of Singapore (NUS) Medical School, Signature Research Program in Cardiovascular and Metabolic Disorders, Singapore, Singapore (GRID:grid.428397.3) (ISNI:0000 0004 0385 0924)
2 Forschungszentrum Jülich GmbH, Institute for Bio- and Geosciences (IBG-5), Jülich, Germany (GRID:grid.8385.6) (ISNI:0000 0001 2297 375X); University of Vienna, Department of Analytical Chemistry, Vienna, Austria (GRID:grid.10420.37) (ISNI:0000 0001 2286 1424)
3 National University of Singapore, Singapore Lipidomics Incubator, Life Sciences Institute, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431)
4 University of Ottawa, 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, Ottawa, Canada (GRID:grid.28046.38) (ISNI:0000 0001 2182 2255)
5 RIKEN Center for Integrative Medical Sciences, Yokohama, Japan (GRID:grid.509459.4) (ISNI:0000 0004 0472 0267); Yokohama City University, Graduate School of Medical Life Science, Yokohama, Japan (GRID:grid.268441.d) (ISNI:0000 0001 1033 6139)
6 Kyushu University, Division of Metabolomics Center, Medical Research Center for High Depth Omics, Medical Institute of Bioregulation, Fukuoka, Japan (GRID:grid.177174.3) (ISNI:0000 0001 2242 4849)
7 MetaboHUBMetaToul Facility, Toulouse, France (GRID:grid.28046.38)
8 Heidelberg University Biochemistry Center (BZH), Heidelberg, Germany (GRID:grid.7700.0) (ISNI:0000 0001 2190 4373)
9 Universidad de los Andes, Metabolomics Core Facility-MetCore, Bogotá, Colombia (GRID:grid.7247.6) (ISNI:0000 0004 1937 0714)
10 Manchester Academic Health Science Centre, 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, United Kingdom (GRID:grid.462482.e) (ISNI:0000 0004 0417 0074)
11 Karolinska Institutet, Unit of Integrative Metabolomics, Institute of Environmental Medicine, Stockholm, Sweden (GRID:grid.4714.6) (ISNI:0000 0004 1937 0626)
12 University of British Columbia, Department of Pathology and Laboratory Medicine, Vancouver, Canada (GRID:grid.17091.3e) (ISNI:0000 0001 2288 9830)
13 University of Pardubice, Department of Analytical Chemistry, Faculty of Chemical Technology, Pardubice, Czech Republic (GRID:grid.11028.3a) (ISNI:0000 0000 9050 662X)
14 Baker Heart and Diabetes Institute, Melbourne, Australia (GRID:grid.1051.5) (ISNI:0000 0000 9760 5620); La Trobe University, Baker Department of Cardiovascular Research, Translation and Implementation, Bundoora, Australia (GRID:grid.1018.8) (ISNI:0000 0001 2342 0938)
15 MetaboHUB, Université Paris-Saclay, CEA, INRAE, Département Médicaments et Technologies pour la Santé (DMTS), Gif sur Yvette, France (GRID:grid.511304.2)
16 University of Vienna, Department of Analytical Chemistry, Vienna, Austria (GRID:grid.10420.37) (ISNI:0000 0001 2286 1424)
17 Avanti Polar Lipids, Alabaster, USA (GRID:grid.423085.8) (ISNI:0000 0004 0595 5711)
18 Babraham Research Campus, Babraham Institute, Cambridge, USA (GRID:grid.423085.8)
19 University of Turku and Åbo Akademi University, Turku Bioscience Centre, Turku, Finland (GRID:grid.1374.1) (ISNI:0000 0001 2097 1371); University of Turku, Department of Chemistry, Turku, Finland (GRID:grid.1374.1) (ISNI:0000 0001 2097 1371)
20 University of Leipzig, Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Leipzig, Germany (GRID:grid.9647.c) (ISNI:0000 0004 7669 9786); University of Leipzig, Center for Biotechnology and Biomedicine, Leipzig, Germany (GRID:grid.9647.c) (ISNI:0000 0004 7669 9786); Faculty of Medicine Carl Gustav Carus of TU Dresden, Center of Membrane Biochemistry and Lipid Research, Dresden, Germany (GRID:grid.4488.0) (ISNI:0000 0001 2111 7257)
21 University of Iceland, Faculty of Pharmaceutical Sciences, Reykjavik, Iceland (GRID:grid.14013.37) (ISNI:0000 0004 0640 0021); ArcticMass, Reykjavik, Iceland (GRID:grid.14013.37)
22 University of Lausanne, Metabolomics Platform, Faculty of Biology and Medicine, Lausanne, Switzerland (GRID:grid.9851.5) (ISNI:0000 0001 2165 4204)
23 Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, The Netherlands (GRID:grid.10419.3d) (ISNI:0000 0000 8945 2978)
24 Nanyang Technological University, Lee Kong Chian School of Medicine, Singapore, Singapore (GRID:grid.59025.3b) (ISNI:0000 0001 2224 0361)
25 German Research Center for Environmental Health, Metabolomics and Proteomics Core, Helmholtz Zentrum München, Neuherberg, Germany (GRID:grid.4567.0) (ISNI:0000 0004 0483 2525)
26 Leiden University, Metabolomics and Analytics Centre, Leiden Academic Centre for Drug Research, Leiden, The Netherlands (GRID:grid.5132.5) (ISNI:0000 0001 2312 1970)
27 Institute of Clinical Chemistry and Laboratory Medicine, University Hospital of Regensburg, Regensburg, Germany (GRID:grid.411941.8) (ISNI:0000 0000 9194 7179)
28 University Zurich, Institute of Clinical Chemistry, Schlieren, Switzerland (GRID:grid.7400.3) (ISNI:0000 0004 1937 0650)
29 Chinese Academy of Sciences, State Key Laboratory of Medical Proteomics, CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Dalian, China (GRID:grid.9227.e) (ISNI:0000000119573309)
30 National Institute of Standards and Technology, Chemical Science Division, Gaithersburg, USA (GRID:grid.94225.38) (ISNI:0000 0004 0506 8207)
31 Medical University of Graz, Core Facility Mass Spectrometry, Graz, Austria (GRID:grid.11598.34) (ISNI:0000 0000 8988 2476)
32 LipidALL Technologies, Changzhou, China (GRID:grid.511275.5); Chinese Academy of Sciences, State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Beijing, China (GRID:grid.9227.e) (ISNI:0000000119573309)
33 University of Leipzig, Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Leipzig, Germany (GRID:grid.9647.c) (ISNI:0000 0004 7669 9786); University of Leipzig, Center for Biotechnology and Biomedicine, Leipzig, Germany (GRID:grid.9647.c) (ISNI:0000 0004 7669 9786)
34 Yonsei University, Department of Chemistry, Seoul, South Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454)
35 Babraham Research Campus, Babraham Institute, Cambridge, USA (GRID:grid.94225.38)
36 Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Centro de Investigaciones en Bionanociencias (CIBION), Ciudad de Buenos Aires, Argentina (GRID:grid.423606.5) (ISNI:0000 0001 1945 2152); Intendente Güiraldes, Departamento de Fisiología, Biología Molecular y Celular, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina (GRID:grid.7345.5) (ISNI:0000 0001 0056 1981); Instituto Nacional de Tecnología Industrial, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Departamento de Desarrollo Analítico y Control de Procesos, Buenos Aires, Argentina (GRID:grid.473334.4) (ISNI:0000 0001 2174 4875)
37 Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Centro de Investigaciones en Bionanociencias (CIBION), Ciudad de Buenos Aires, Argentina (GRID:grid.423606.5) (ISNI:0000 0001 1945 2152); Departamento de Ciencias Químicas, Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Junin, Argentina (GRID:grid.7345.5) (ISNI:0000 0001 0056 1981)
38 Tata Institute of Fundamental Research, National Centre for Biological Sciences, Bangalore, India (GRID:grid.22401.35) (ISNI:0000 0004 0502 9283)
39 Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Centro de Investigaciones en Bionanociencias (CIBION), Ciudad de Buenos Aires, Argentina (GRID:grid.423606.5) (ISNI:0000 0001 1945 2152)
40 National University of Singapore, Singapore Lipidomics Incubator, Life Sciences Institute, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431); Tata Institute of Fundamental Research, National Centre for Biological Sciences, Bangalore, India (GRID:grid.22401.35) (ISNI:0000 0004 0502 9283)
41 Manchester Academic Health Science Centre, 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, United Kingdom (GRID:grid.462482.e) (ISNI:0000 0004 0417 0074); Manchester Academic Health Science Centre, Lydia Becker Institute of Immunology and Inflammation; Faculty of Biology, Medicine and Health, The University of Manchester, Manchester, United Kingdom (GRID:grid.462482.e) (ISNI:0000 0004 0417 0074)
42 Cardiff University, Systems Immunity Research Institute, School of Medicine, Cardiff, UK (GRID:grid.5600.3) (ISNI:0000 0001 0807 5670)
43 University of Turku and Åbo Akademi University, Turku Bioscience Centre, Turku, Finland (GRID:grid.1374.1) (ISNI:0000 0001 2097 1371); Örebro University, School of Medical Sciences, Faculty of Medicine and Health, Örebro, Sweden (GRID:grid.15895.30) (ISNI:0000 0001 0738 8966)
44 Institute for Stem Cell Science and Regenerative Medicine, Bangalore, India (GRID:grid.475408.a) (ISNI:0000 0004 4905 7710)
45 Teikyo University, Laboratory of Biomedical and Analytical Sciences, Faculty of Pharma-Science, Itabashi-ku, Japan (GRID:grid.264706.1) (ISNI:0000 0000 9239 9995); Tohoku University, Department of Integrative Genomics, Tohoku Medical Megabank Organization, Sendai, Japan (GRID:grid.69566.3a) (ISNI:0000 0001 2248 6943)
46 National Institute of Standards and Technology, Chemical Science Division, Charleston, USA (GRID:grid.94225.38) (ISNI:0000 0004 0506 8207)
47 Vienna (BOKU), Christian Doppler Laboratory for Innovative Gut Health Concepts of Livestock, Institute of Bioanalytics and Agro-Metabolomics, Department of Agrobiotechnology (IFATulln), University of Natural Resources and Life Sciences, Tulln, Austria (GRID:grid.5173.0) (ISNI:0000 0001 2298 5320)
48 Chinese Academy of Sciences, State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Beijing, China (GRID:grid.9227.e) (ISNI:0000000119573309)
49 MetaboHUBMetaToul Facility, Toulouse, France (GRID:grid.177174.3)
50 RIKEN Center for Integrative Medical Sciences, Yokohama, Japan (GRID:grid.509459.4) (ISNI:0000 0004 0472 0267); Yokohama City University, Graduate School of Medical Life Science, Yokohama, Japan (GRID:grid.268441.d) (ISNI:0000 0001 1033 6139); RIKEN Center for Sustainable Resource Science, Yokohama, Japan (GRID:grid.509461.f) (ISNI:0000 0004 1757 8255); Tokyo University of Agriculture and Technology, Department of Biotechnology and Life Science, Tokyo, Japan (GRID:grid.136594.c) (ISNI:0000 0001 0689 5974)
51 Department of Pathology and Laboratory Medicine, St. Paul’s Hospital, Vancouver, Canada (GRID:grid.416553.0) (ISNI:0000 0000 8589 2327)
52 Babraham Research Campus, Babraham Institute, Cambridge, USA (GRID:grid.416553.0)
53 Karolinska Institutet, Unit of Integrative Metabolomics, Institute of Environmental Medicine, Stockholm, Sweden (GRID:grid.4714.6) (ISNI:0000 0004 1937 0626); Karolinska University Hospital, Department of Respiratory Medicine and Allergy, Stockholm, Sweden (GRID:grid.24381.3c) (ISNI:0000 0000 9241 5705)
54 University of Florida, Department of Physiological Sciences, College of Veterinary Medicine, Gainesville, USA (GRID:grid.15276.37) (ISNI:0000 0004 1936 8091)
55 Lipidomics Consulting Ltd., Espoo, Finland (GRID:grid.519011.d)
56 National University of Singapore, Singapore Lipidomics Incubator, Life Sciences Institute, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431); National University of Singapore, Precision Medicine Translational Research Programme and Department of Biochemistry, Yong Loo Lin School of Medicine, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431); Hamad Bin Khalifa University, College of Health and Life Sciences, Doha, Qatar (GRID:grid.452146.0) (ISNI:0000 0004 1789 3191)