U.S. Department of Energy

Pacific Northwest National Laboratory

A reversed-phase capillary ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for comprehensive top-down/bottom-up lipid profiling.

TitleA reversed-phase capillary ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for comprehensive top-down/bottom-up lipid profiling.
Publication TypeJournal Article
Year of Publication2012
AuthorsGao X, Zhang Q, Meng D, Isaac G, Zhao R, Fillmore TL, Chu RK, Zhou J, Tang K, Hu Z, Moore RJ, Smith RD, Katze MG, Metz TO
JournalAnal Bioanal Chem
KeywordsAnimals, Cell Line, Chromatography, High Pressure Liquid, Chromatography, Reverse-Phase, Lipids, Mass Spectrometry, Rats
Abstract

Lipidomics is a critical part of metabolomics and aims to study all the lipids within a living system. We present here the development and evaluation of a sensitive capillary UPLC-MS method for comprehensive top-down/bottom-up lipid profiling. Three different stationary phases were evaluated in terms of peak capacity, linearity, reproducibility, and limit of quantification (LOQ) using a mixture of lipid standards representative of the lipidome. The relative standard deviations of the retention times and peak abundances of the lipid standards were 0.29% and 7.7%, respectively, when using the optimized method. The linearity was acceptable at >0.99 over 3 orders of magnitude, and the LOQs were sub-fmol. To demonstrate the performance of the method in the analysis of complex samples, we analyzed lipids extracted from a human cell line, rat plasma, and a model human skin tissue, identifying 446, 444, and 370 unique lipids, respectively. Overall, the method provided either higher coverage of the lipidome, greater measurement sensitivity, or both, when compared to other approaches of global, untargeted lipid profiling based on chromatography coupled with MS.

DOI10.1007/s00216-012-5773-5
Alternate JournalAnal Bioanal Chem
PubMed ID22354571
PubMed Central IDPMC3531553
Grant ListHHSN272200800060C / / PHS HHS / United States
U54 AI081680 / AI / NIAID NIH HHS / United States
U54AI081680 / AI / NIAID NIH HHS / United States
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