An Interlaboratory Comparison of High-Resolution Ion Mobility Collision Cross-Sections from Human Plasma

The increasing adoption of high-resolution ion mobility (HRIM) in untargeted omics workflows underscores the need for precise collision cross-section (CCS) measurements which are highly reproducible across various laboratories and instrumentation. To evaluate the reproducibility of a high-resolution ion mobility platform, an interlaboratory study was undertaken using structures for lossless ion manipulation-based traveling wave ion mobility spectrometry (TWSLIM) in nitrogen drift gas. Across 250 lipid features spanning glycerophospholipids, glycerolipids, and sphingolipids detected from a lipidomic extract of human plasma standard reference material, the platform demonstrated high CCS measurement reproducibility, with an average relative standard deviation (%RSD) of ∼0.1%. Triglycerides in general were found to exhibit multiple IM features that served as an illustrative example where analysis via HRIM and data-independent, mobility-aligned fragmentation (MAF) provides critical insights into their chemical structures. To support the future development of HRIM in lipidomic workflows, a large (n = 250) number of lipid consensus ^TWSLIMCCS_N2 values was compiled from the interlaboratory study into a HRIM database for community use.