Journal
ANALYTICAL CHEMISTRY
Volume 89, Issue 1, Pages 656-665Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b02930
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Funding
- FP7 project, EuroMotor [259867]
- FP7 project, HECATOS [602156]
- FP7 project, HELIX [603864]
- FP7 project, DETECTIVE [266838]
- Medical Research Council [MC_UP_A090_1006, MC_PC_13030, MC_PC_13046]
- BBSRC [BB/M027252/1, BB/I019405/1, BB/M027252/2] Funding Source: UKRI
- MRC [MR/P011705/1, MC_PC_13030, MC_UP_A090_1006, MR/P01836X/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/M027252/2, BB/M027252/1] Funding Source: researchfish
- Cancer Research UK [11566] Funding Source: researchfish
- Medical Research Council [MR/P01836X/1, MC_PC_13046, MC_PC_13030, MC_UP_A090_1006, MR/P011705/1] Funding Source: researchfish
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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 AbsolutelDQ 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 F1A measurements. This is the first interlahotatory 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.
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