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

Xiaoli Gao, Qibin Zhang, Da Meng, Giorgis Isaac, Rui Zhao, Thomas L. Fillmore, Rosey K. Chu, Jianying Zhou, Keqi Tang, Zeping Hu, Ronald J. Moore, Richard D. Smith, Michael G. Katze, Thomas O. Metz

Research output: Contribution to journalArticle

73 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2923-2933
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
Volume402
Issue number9
DOIs
StatePublished - Mar 1 2012

Keywords

  • Electrospray ionization (ESI)
  • Tandem mass spectrometry (MS/MS)
  • Top-down/bottom-up lipid profiling
  • Ultra-performance liquid chromatography (UPLC)

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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    Gao, X., Zhang, Q., Meng, D., Isaac, G., Zhao, R., Fillmore, T. L., Chu, R. K., Zhou, J., Tang, K., Hu, Z., Moore, R. J., Smith, R. D., Katze, M. G., & Metz, T. O. (2012). A reversed-phase capillary ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for comprehensive top-down/bottom-up lipid profiling. Analytical and Bioanalytical Chemistry, 402(9), 2923-2933. https://doi.org/10.1007/s00216-012-5773-5