title: Profiling and relative quantification of phosphatidylethanolamine based on acetone stable isotope derivatization.
abstract: Phosphatidylethanolamine (PE) is considered to be one of the pivotal lipids for normal cellular function as well as disease initiation and progression. In this study, a simple, efficient, reliable, and inexpensive method for the qualitative analysis and relative quantification of PE, based on acetone stable isotope derivatization combined with double neutral loss scan-shotgun electrospray ionization tandem-quadrupole mass spectrometry analysis (ASID-DNLS-Shotgun ESI-MS/MS), was developed. The ASID method led to alkylation of the primary amino groups of PE with an isopropyl moiety. The use of acetone (d0-acetone) and deuterium-labeled acetone (d6-acetone) introduced a 6 Da mass shift that was ideally suited for relative quantitative analysis, and enhanced sensitivity for mass analysis. The DNLS model was introduced to simultaneously analyze the differential derivatized PEs by shotgun ESI-MS/MS with high selectivity and accuracy. The reaction specificity, labeling efficiency, and linearity of the ASID method were thoroughly evaluated in this study. Its excellent applicability was validated by qualitative and relative quantitative analysis of PE species presented in liver samples from rats fed different diets. Using the ASID-DNLS-Shotgun ESI-MS/MS method, 45 PE species from rat livers have been identified and quantified in an efficient manner. The level of total PEs tended to decrease in the livers of rats on high fat diets compared with controls. The levels of PE 32:1, 34:3, 34:2, 36:3, 36:2, 42:10, plasmalogen PE 36:1 and lyso PE 22:6 were significantly reduced, while levels of PE 36:1 and lyso PE 16:0 increased.
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, PR China; Hubei Key Laboratory of Lipid Chemistry and Nutrition, PR China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, PR China; Hubei Key Laboratory of Lipid Chemistry and Nutrition, PR China. Electronic address: willasa@163.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Wei
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, PR China; Hubei Key Laboratory of Lipid Chemistry and Nutrition, PR China.
authors.forename: Ji-Qu
authors.initials: JQ
authors.lastname: Xu
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, PR China; Hubei Key Laboratory of Lipid Chemistry and Nutrition, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Lv
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, PR China; Hubei Key Laboratory of Lipid Chemistry and Nutrition, PR China.
authors.forename: Xu-Yan
authors.initials: XY
authors.lastname: Dong
authors.affiliation: Center for Metabolic Origins of Disease, Sanford Burnham Prebys Medical Discovery Institute, Orlando, FL 32827, USA; College of Basic Medical Sciences, Zhejiang Chinese Medical University, 548 Bingwen Road, Hangzhou, Zhejiang 310053, PR China.
authors.forename: Xianlin
authors.initials: X
authors.lastname: Han
authors.affiliation: School of Chemical Science, The University of Auckland, Auckland 1142, New Zealand.
authors.forename: Siew-Young
authors.initials: SY
authors.lastname: Quek
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, PR China; Hubei Key Laboratory of Lipid Chemistry and Nutrition, PR China.
authors.forename: Feng-Hong
authors.initials: FH
authors.lastname: Huang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, PR China; Hubei Key Laboratory of Lipid Chemistry and Nutrition, PR China. Electronic address: chenhong@oilcrops.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
mesh_terms: D000096:Acetone / Q000737:chemistry*; D000818:Animals; D007554:Isotopes / Q000737:chemistry*; D057230:Limit of Detection; D008099:Liver / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D051381:Rats
chemical_list: D007554:Isotopes; D010714:Phosphatidylethanolamines; D000096:Acetone; C483858:phosphatidylethanolamine
keywords: Acetone stable isotope derivatization; Mass spectrometry; Phosphatidylethanolamine; Relative quantification