Targeted Metabolomic Approach to Assess the Reproducibility of Plasma Metabolites over a Four Month Period in a Free-Living Population

[Image: see text] Metabolomics is increasingly applied to investigate diet–disease associations in nutrition research. However, studies of metabolite reproducibility are limited, which could hamper their use within epidemiologic studies. The objective of this study was to evaluate the metabolite reproducibility during 4 months in a free-living population. In the A-DIET Confirm study, fasting plasma and dietary data were collected once a month for 4 months. Metabolites were measured using liquid chromatography tandem mass spectrometry, and their reproducibility was estimated using the intraclass correlation coefficient (ICC). Regularized canonical correlation analysis (rCCA) was employed to examine the diet–metabolite associations. In total, 138 metabolites were measured, and median ICC values of 0.49 and 0.65 were found for amino acids and biogenic amines, respectively. Acylcarnitines, lysophosphatidylcholines, phosphatidylcholines, and sphingomyelins had median ICC values of 0.69, 0.66, 0.63, and 0.63, respectively. The median ICC for all metabolites was 0.62, and 54% of metabolites had ICC values ≥0.60. Additionally, the rCCA heat map revealed positive correlations between dairy/meat intake and specific lipids. In conclusion, more than half of the metabolites demonstrated good to excellent reproducibility. A single measurement per subject could appropriately reflect the metabolites’ long-term concentration levels and may also be sufficient for assessing disease risk in epidemiologic studies. The study data are deposited in MetaboLights (MTBLS3428 (www.ebi.ac.uk/metabolights)).