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

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 the 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 they 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. Interlaboratory 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.