International Ring Trial of a High Resolution Targeted Metabolomics and Lipidomics Platform for Serum and Plasma Analysis

Abstract

A challenge facing metabolomics in the analysis of large human cohorts is the cross-laboratory comparability of quantitative metabolomics measurements. In this study, 14 laboratories analyzed various blood specimens using a common experimental protocol provided with Biocrates AbsoluteIDQ p400HR kit, to quantify up to 408 metabolites. The specimens included human plasma and serum from male and female donors, mouse and rat plasma as well as NIST SRM 1950 reference plasma. The metabolite classes covered range from polar (e.g. amino acids and biogenic amines), to nonpolar (e.g. diacyl-and triacyl-glycerols), and span 11 common metabolite classes. The manuscript describes a strict system suitability testing (SST) criteria used to evaluate each laboratory's readiness to perform the assay, and provides the SST Skyline documents for public dissemination. The study found approximately 250 metabolites were routinely quantified in the sample types tested, using Orbitrap instruments. Inter-laboratory variance for the NIST SRM-1950 has a median of 10% for amino acids, 24% for biogenic amines, 38% for acylcarnitines, 25% for glycerolipids, 23% for glycerophospholipids, 16% for cholesteryl esters, 15% for sphingolipids, and 9% for hexoses. Comparing to consensus values for NIST SRM-1950, nearly 80% of comparable analytes demonstrated bias of <50% from the reference value. The findings of this study result in recommendations of best practices for system suitability, quality control, and calibration. We demonstrate that with appropriate controls, high-resolution metabolomics can provide accurate results with good precision across laboratories, and the p400HR therefore is a reliable approach for generating consistent and comparable metabolomics data.

Nrf2 contributes to the weight gain of mice during space travel

“…The ultra-high-performance liquid chromatography (UHPLC) system consisted of an online degasser, auto sampler, dual pump, and column oven (UltiMate TM 3000 RSLC system, Thermo Fisher Scientific), and a quadrupole Fourier transform mass spectrometry (FTMS, Q Exactive Orbitrap system). The operating conditions of the UHPLC-FTMS system and the details of data analysis followed the protocol of the Kit400 analysis 43 .…”

A comparative evaluation of the generalised predictive ability of eight machine learning algorithms across ten clinical metabolomics data sets for binary classification

“…Most large academic laboratories also provide some level of similar service. This is illustrated by the recent successful ring trial for the Biocrates AbsoluteIDQ p400HR assay (Thompson et al 2019) which will allow data sets from multiple labs to be potentially combined into one data analysis. Other approaches to data fusion have most recently been reported in the American Journal of Epidemiology by Yu et al (2019) “Consortium of Metabolomics Studies (COMETS) Metabolomics in 47 Prospective Cohort Studies”.…”

A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with13C- internal standards

“…[40][41][42] Human plasma lipidomics serves as prime example to show the validity of the presented workflow as it represents the most frequently analyzed sample matrix in the field. 1,24 The presented validation capitalizes on healthy donor samples [43][44][45][46][47] and reference materials 46 with published quantitative lipidomes. However, a certified reference material ensuring traceability is still lacking in the field.…”