Two‐dimensional LC‐MS/MS to enhance ceramide and phosphatidylcholine species profiling in mouse liver |
| |
| Authors: | Yee Soon Ling Hao‐Jan Liang Ming‐Huan Lin Chuan‐Ho Tang Kuen‐Yuh Wu Min‐Liang Kuo Ching Yu Lin |
| |
| Institution: | 1. Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan;2. Institute of Environmental Health, College of Public Health, National Taiwan University, Taipei, Taiwan;3. Institute of Occupational Medicine and Industrial Hygiene, College of Public Health, National Taiwan University, Taipei, Taiwan;4. National Museum of Marine Biology and Aquarium, Pingtung, Taiwan;5. Institute of Marine Biodiversity and Evolutionary Biology, National Dong Hwa University, Pingtung, Taiwan;6. College of Life Science, National Taiwan University, Taipei, Taiwan |
| |
| Abstract: | 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 system used a silica HILIC column operated in the first dimension to distinguish the lipid classes and a BEH C18 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 © 2014 John Wiley & Sons, Ltd. |
| |
| Keywords: | 2D LC‐MS/MS lipidomics ceramide phosphatidylcholine mouse liver |
|
|
