A significant challenge facing the vision of metabolomics in precision medicine, and the analysis of large human cohorts, is the cross-laboratory and longitudinal comparability of quantitative measurements and laboratory protocols used. In this study, 14 laboratories world-wide 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 targets covered from very polar (amino acids and biogenic amines), to very nonpolar (diacyl- and triacyl-glycerols), and spanned across 11 common metabolite classes. The manuscript describes a strict system suitability testing (SST) criteria used to first evaluate each laboratory’s readiness to perform the assay, and provides the SST Skyline documents for public dissemination. Approximately 250 metabolites were routinely quantified with the kit using various models of high-resolution 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. This study provides recommendations for large metabolomics studies in terms of best practices for system suitability, quality control, and calibration; and has demonstrated that high-resolution metabolomics methods provide excellent quantitative precision across laboratories.

This documentation includes two Skyline files, one for the UHPLC-MS measurement of amino acids and biogenic amines using the kits, and a second using flow-injeciton analysis (FIA)-MS for the purposes of testing measurement capability for acylcarnitines, hexoses, and lipids. The SST solution is made up of a mix of stable-isotope labeled standards.

A mix of neat standards distributed with each kit for the purposes of establishing System Suitability prior to attempting the p400HR analysis.