title: Interlaboratory Reproducibility of a Targeted Metabolomics Platform for Analysis of Human Serum and Plasma.
abstract: A critical question facing the field of metabolomics is whether data obtained from different centers can be effectively compared and combined. An important aspect of this is the interlaboratory precision (reproducibility) of the analytical protocols used. We analyzed human samples in six laboratories using different instrumentation but a common protocol (the AbsoluteIDQ p180 kit) for the measurement of 189 metabolites via liquid chromatography (LC) or flow injection analysis (FIA) coupled to tandem mass spectrometry (MS/MS). In spiked quality control (QC) samples 82% of metabolite measurements had an interlaboratory precision of <20%, while 83% of averaged individual laboratory measurements were accurate to within 20%. For 20 typical biological samples (serum and plasma from healthy individuals) the median interlaboratory coefficient of variation (CV) was 7.6%, with 85% of metabolites exhibiting a median interlaboratory CV of <20%. Precision was largely independent of the type of sample (serum or plasma) or the anticoagulant used but was reduced in a sample from a patient with dyslipidaemia. The median interlaboratory accuracy and precision of the assay for standard reference plasma (NIST SRM 1950) were 107% and 6.7%, respectively. Likely sources of irreproducibility were the near limit of detection (LOD) typical abundance of some metabolites and the degree of manual review and optimization of peak integration in the LC-MS/MS data after acquisition. Normalization to a reference material was crucial for the semi-quantitative FIA measurements. This is the first interlaboratory assessment of a widely used, targeted metabolomics assay illustrating the reproducibility of the protocol and how data generated on different instruments could be directly integrated in large-scale epidemiological studies.
authors.affiliation: Department of Surgery and Cancer, Imperial College London , London, W12 0NN, U.K.
authors.forename: Alexandros P
authors.identifier: 0000-0002-5635-7426
authors.initials: AP
authors.lastname: Siskos
authors.affiliation: Department of Surgery and Cancer, Imperial College London , London, W12 0NN, U.K.
authors.forename: Pooja
authors.initials: P
authors.lastname: Jain
authors.affiliation: Institute of Bioinformatics and Systems Biology, Helmholtz Zentrum München, German Research Center for Environmental Health , 85764 Neuherberg, Germany.
authors.forename: Werner
authors.initials: W
authors.lastname: Römisch-Margl
authors.affiliation: Department of Life Sciences, Imperial College London , London, SW7 2AZ, U.K.
authors.forename: Mark
authors.initials: M
authors.lastname: Bennett
authors.affiliation: International Agency for Research on Cancer (IARC) , Nutrition and Metabolism Section, Biomarkers Group, F-69372 Lyon, Cedex 08, France.
authors.forename: David
authors.initials: D
authors.lastname: Achaintre
authors.affiliation: The Institute of Cancer Research , Sutton, SM2 5NG, U.K.
authors.forename: Yasmin
authors.initials: Y
authors.lastname: Asad
authors.affiliation: MRC Human Nutrition Research , Cambridge, CB1 9NL, U.K.
authors.forename: Luke
authors.initials: L
authors.lastname: Marney
authors.affiliation: MRC Human Nutrition Research , Cambridge, CB1 9NL, U.K.
authors.forename: Larissa
authors.initials: L
authors.lastname: Richardson
authors.affiliation: MRC Human Nutrition Research , Cambridge, CB1 9NL, U.K.
authors.forename: Albert
authors.initials: A
authors.lastname: Koulman
authors.affiliation: MRC Human Nutrition Research , Cambridge, CB1 9NL, U.K.
authors.forename: Julian L
authors.initials: JL
authors.lastname: Griffin
authors.affiliation: The Institute of Cancer Research , Sutton, SM2 5NG, U.K.
authors.forename: Florence
authors.initials: F
authors.lastname: Raynaud
authors.affiliation: International Agency for Research on Cancer (IARC) , Nutrition and Metabolism Section, Biomarkers Group, F-69372 Lyon, Cedex 08, France.
authors.forename: Augustin
authors.initials: A
authors.lastname: Scalbert
authors.affiliation: Genome Analysis Center, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health , 85764 Neuherberg, Germany.|Chair of Experimental Genetics, Center of Life and Food Sciences Weihenstephan, Technische Universität München , 85354 Freising-Weihenstephan, Germany.|German Center for Diabetes Research , 85764 Neuherberg, Germany.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Adamski
authors.affiliation: Genome Analysis Center, Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health , 85764 Neuherberg, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Prehn
authors.affiliation: Department of Surgery and Cancer, Imperial College London , London, W12 0NN, U.K.
authors.forename: Hector C
authors.initials: HC
authors.lastname: Keun
mesh_terms: D006801:Humans; D007753:Laboratories; D057230:Limit of Detection; D055432:Metabolomics / Q000379:methods* / Q000592:standards; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry