title: Two-dimensional LC-MS/MS to enhance ceramide and phosphatidylcholine species profiling in mouse liver.
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.
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, No. 1, Sec. 1, Jen-ai Rd, Taipei, 100, Taiwan; Institute of Environmental Health, College of Public Health, National Taiwan University, No. 17, Xu-zhou Rd, Taipei, 100, Taiwan.
authors.forename: Yee Soon
authors.initials: YS
authors.lastname: Ling
authors.forename: Hao-Jan
authors.initials: HJ
authors.lastname: Liang
authors.forename: Ming-Huan
authors.initials: MH
authors.lastname: Lin
authors.forename: Chuan-Ho
authors.initials: CH
authors.lastname: Tang
authors.forename: Kuen-Yuh
authors.initials: KY
authors.lastname: Wu
authors.forename: Min-Liang
authors.initials: ML
authors.lastname: Kuo
authors.forename: Ching Yu
authors.initials: CY
authors.lastname: Lin
mesh_terms: D000818:Animals; D002518:Ceramides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D057230:Limit of Detection; D016014:Linear Models; D008099:Liver / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D055432:Metabolomics; D051379:Mice; D008813:Mice, Inbred ICR; D009281:Naphthalenes / Q000633:toxicity; D010713:Phosphatidylcholines / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002518:Ceramides; D009281:Naphthalenes; D010713:Phosphatidylcholines; C031721:naphthalene
keywords: 2D LC-MS/MS; ceramide; lipidomics; mouse liver; phosphatidylcholine