A Real-Life Reproducibility Assessment for NMR Metabolomics

Abstract

Nuclear magnetic resonance (NMR) metabolomics is currently popular enough to attract both specialized and non-specialized NMR groups involving both analytical trained personnel and newcomers, including undergraduate students. Recent interlaboratory studies performed by established NMR metabolomics groups demonstrated high reproducibility of the state-of-the-art NMR equipment and SOPs. There is, however, no assessment of NMR reproducibility when mixing both analytical experts and newcomers. An interlaboratory assessment of NMR quantitation reproducibility was performed using two NMR instruments belonging to different laboratories and involving several operators with different backgrounds and metabolomics expertise for the purpose of assessing the limiting factors for data reproducibility in a multipurpose NMR environment. The variability induced by the operator, automatic pipettes, NMR tubes and NMR instruments was evaluated in order to assess the limiting factors for quantitation reproducibility. The results estimated the expected reproducibility data in a real-life multipurpose NMR laboratory to a maximum 4% variability, demonstrating that the current NMR equipment and SOPs may compensate some of the operator-induced variability.

Lipoprofiling Assessed by NMR Spectroscopy in Patients with Acute Coronary Syndromes: Is There a Need for Fasting Prior to Sampling?

“…On the contrary, for 56 out of the total of 68 tests, more than one serum vial was analyzed separately, and the results of these multiple analyses were averaged in order to improve the analytical accuracy. The accuracy and reproducibility of NMR metabolomics and lipidomics has been recently demonstrated by large trials [ 8 , 9 , 11 , 12 ], and we have also described recent reproducibility assessments for the NMR method [ 7 , 13 ]. A subset of samples was checked for some parameters (total cholesterol, total triglycerides, total LDL, and total HDL) through both NMR standardized methods of the hospital’s biochemistry laboratory, and the two methods are in good agreement ( Supplementary Table S2 ).…”

“…A volume of 600 µL of the homogenized sample was then transferred into a 5 mm NMR tube (Wilmad 507) and further loaded into the NMR sample changer. The operator’s performance and the NMR tubes’ quality were assessed as we previously described [ 7 ]. The NMR experiments were performed with an Avance III HD 600 NMR spectrometer (Bruker Biospin, Ettlingen) operating at 600.12 MHz for 1 H nuclei, equipped with a 5 mm inverse detection (BBI) probe with gradients on the z -axis.…”

Inter-laboratory comparison of plant volatile analyses in the light of intra-specific chemodiversity

“…In recent years, the reproducibility of scientific results has been called into question (Ioannidis, 2005) and this problem has also been acknowledged in metabolomics studies (Ghosh et al, 2021; Stavarache et al, 2022). Using a comparative inter-laboratory ring trial, we aimed at identifying sources of variation that may influence the reproducibility of obtained results on plant VOC profiles when applying the PDMS approach coupled with TD-GC-MS analysis in collaborative projects.…”

Inter-laboratory comparison of plant volatile analyses in the light of intra-specific chemodiversity

“…In recent years, the reproducibility of scientific results has been called into question (Ioannidis, 2005 ) and this problem has also been acknowledged in metabolomics studies (Ghosh et al, 2021 ; Stavarache et al, 2022 ). Using a comparative inter-laboratory ring trial, we aimed at identifying sources of variation that may influence the reproducibility of obtained results on plant VOC profiles when applying the PDMS approach coupled with TD-GC-MS analysis in collaborative projects.…”