Enrichment of resolving power improves ion-peak quantification on a lipidomics platform.

In this study, we delineated the importance of MS resolving power on the ion-peak quantification of lipids using an Orbitrap Fusion instrument and established a liquid chromatography-based, high-performance lipidomics platform. The ion-peak recognition of several lipids in human plasma, such as LPC(15:0), LPE(22:5), and PC(35:0), was clearly improved by increasing the MS resolving power. In addition, we evaluated the impact of resolving power on the quantitative detection of lipids by automatic ion-peak recognition with calculation of the coefficient of variance (CV). The extracted ions obtained from human plasma were automatically annotated by Compound Discoverer software with manual confirmation of standards or MS/MS fragments (class- and acyl side chain-specific ions and neutral losses). Quantitative evaluation of 499 lipids in human plasma in terms of their CV values clearly demonstrated an improvement in the quantitative performance by enriching the resolving power. Moreover, we evaluated our new lipidomics platform with enriched MS resolving power (setting of 240,000, full width at half maximum at m/z 200). Because automatic annotation by TraceFinder software overlooks several lipid ions, we further manually annotated additional lipid ions, which were confirmed by standards or MS/MS fragments. Eventually, our platform detected 967 lipids encompassing 34 lipid classes, which were confirmed with standards or MS/MS fragments. Of these lipids, 922 scored <20% of the CV values. Taken together, enriching the resolving power improved ion-peak quantification on our novel lipidomics platform, which enabled us to detect broad-spectrum lipids from human plasma.