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Interlaboratory Reproducibility of a Targeted Metabolomics Platform for Analysis of Human Serum and Plasma.

Accepted version
Peer-reviewed

Type

Article

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Authors

Siskos, Alexandros P  ORCID logo  https://orcid.org/0000-0002-5635-7426
Jain, Pooja 
Römisch-Margl, Werner 
Bennett, Mark 
Achaintre, David 

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.

Description

Keywords

Humans, Laboratories, Limit of Detection, Metabolomics, Quality Control, Reference Standards, Reproducibility of Results, Tandem Mass Spectrometry

Journal Title

Anal Chem

Conference Name

Journal ISSN

0003-2700
1520-6882

Volume Title

89

Publisher

American Chemical Society (ACS)
Sponsorship
Medical Research Council (MR/P011705/1)
Medical Research Council (MR/P01836X/1)
Biotechnology and Biological Sciences Research Council (BB/M027252/2)
Medical Research Council (MC_PC_13030)
Biotechnology and Biological Sciences Research Council (BB/M027252/1)
Medical Research Council (MC_PC_13046)