TY - JOUR
T1 - A reversed-phase capillary ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for comprehensive top-down/bottom-up lipid profiling
AU - Gao, Xiaoli
AU - Zhang, Qibin
AU - Meng, Da
AU - Isaac, Giorgis
AU - Zhao, Rui
AU - Fillmore, Thomas L.
AU - Chu, Rosey K.
AU - Zhou, Jianying
AU - Tang, Keqi
AU - Hu, Zeping
AU - Moore, Ronald J.
AU - Smith, Richard D.
AU - Katze, Michael G.
AU - Metz, Thomas O.
N1 - Funding Information:
Acknowledgments This work was supported by the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health, Department of Health and Human Services, under Contract Number HHSN272200800060C. Additional support was provided by the NIAID under Award Number U54AI081680 and by the Office of Science, U.S. Department of Energy (DOE), under the Low Dose Radiation Research Program. Work was performed at the Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by the DOE’s Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory (PNNL) in Richland, Washington. PNNL is a multi-program national laboratory operated by Battelle for the DOE under Contract DE-AC05-76RLO 1830.
PY - 2012/3
Y1 - 2012/3
N2 - 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.
AB - 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.
KW - Electrospray ionization (ESI)
KW - Tandem mass spectrometry (MS/MS)
KW - Top-down/bottom-up lipid profiling
KW - Ultra-performance liquid chromatography (UPLC)
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U2 - 10.1007/s00216-012-5773-5
DO - 10.1007/s00216-012-5773-5
M3 - Article
C2 - 22354571
AN - SCOPUS:84862778693
SN - 1618-2642
VL - 402
SP - 2923
EP - 2933
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 9
ER -