Journal
BIOMEDICAL CHROMATOGRAPHY
Volume 28, Issue 9, Pages 1284-1293Publisher
WILEY
DOI: 10.1002/bmc.3162
Keywords
2D LC-MS/MS; lipidomics; ceramide; phosphatidylcholine; mouse liver
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Funding
- National Science Council [NSC 99-2314-B-002-127-MY3]
- National Health Research Institutes of Taiwan [NHRI-EX100-9915EC, NHRI-EX101-9915EC, EX102-9915EC]
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A two-dimensional (2D) hydrophilic interaction liquid chromatography (HILIC) and reverse-phase (RP) liquid chromatography (LC) system coupled with triple-quadrupole mass spectrometry (MS) was developed to comprehensively profile ceramides and phosphatidylcholine in extracted biological samples. Briefly, the 2D HILIC-RPLC systemused a silica HILIC column operated in the first dimension to distinguish the lipid classes and a BEH C-18 column operated in the second dimension to separate the lipid species of the same class. The regression linearity of each lipid was satisfactory in both systems; however, the absolute matrix effect factor was reduced in 2D LC-MS/MS system. Limits of detection of 2D LC-MS/MS system were 2- to 3-fold lower compared with one-dimensional RPLC-MS/MS. The recovery from the sample ranged from 84.5 to 110%. To summarize, the developed method was proven to be accurate and producible, as relative standard deviations remained <20% at three spiked levels. The efficiency of this newly developed system was applied to measure changes of lipids in the liver of mice after naphthalene treatment. Orthogonal projection to latent structures-discriminant analysis discriminated the lipids from control and the treatment group. We concluded that 2D LC-MS/MS is a promising method to assist lipidomic studies of complex biological samples. Copyright (C) 2014 John Wiley & Sons, Ltd.
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