title: High-throughput and rapid quantification of lipids by nanoflow UPLC-ESI-MS/MS: application to the hepatic lipids of rabbits with nonalcoholic fatty liver disease.
abstract: A rapid and high-throughput quantification method (approximately 300 lipids within 20 min) was established using nanoflow ultrahigh-pressure liquid chromatography-tandem mass spectrometry (nUPLC-ESI-MS/MS) with selective reaction monitoring (SRM) and applied to the quantitative profiling of the hepatic lipids of rabbits with different metabolic conditions that stimulate the development of nonalcoholic fatty liver disease (NAFLD). Among the metabolic conditions of rabbits in this study [inflammation (I), high-cholesterol diet (HC), and high-cholesterol diet combined with inflammation (HCI)], significant perturbation in hepatic lipidome (>3-fold and p < 0.01) was observed in the HC and HCI groups, while no single lipid showed a significant change in group I. In addition, this study revealed a dramatic increase (>2-fold) in relatively high-abundant monohexosylceramides (MHCs), sphingomyelins (SMs), and triacylglycerols (TGs) in both the HC and HCI groups, especially in MHCs as all 11 MHCs increased by larger than 3- to 12-fold. As the levels of the relatively high-abundant lipids in the above classes increased, the total lipidome level of each class increased significantly by approximately 2-fold to 5-fold. Other classes of lipids also generally increased, which was likely induced by the increase in mitogenic and nonapoptotic MHCs and SMs, as they promote cell proliferation. On the other hand, a slight decrease in the level of apoptotic ceramides (Cers) was observed, which agreed with the general increase in total lipid level. As distinct changes in hepatic lipidome were observed from HC groups, this suggests that HC or HCI is highly associated with NAFLD but not inflammation alone itself. Graphical Abstract Schematic of lipidomic analysis from hepatic tissue using nanoflow LC-ESI-MS/MS and nUPLC-ESI-MS/MS.
authors.affiliation: Department of Chemistry, Yonsei University, Seoul, 03722, Republic of Korea.
authors.forename: Seul Kee
authors.initials: SK
authors.lastname: Byeon
authors.affiliation: Department of Chemistry, Yonsei University, Seoul, 03722, Republic of Korea.
authors.forename: Jong Cheol
authors.initials: JC
authors.lastname: Lee
authors.affiliation: Future Convergence Research Division, Korea Institute of Science and Technology, Seoul, 02792, Republic of Korea.
authors.forename: Bong Chul
authors.initials: BC
authors.lastname: Chung
authors.affiliation: Cardiovascular Center, Korea University Guro Hospital, Seoul, 08308, Republic of Korea.
authors.forename: Hong Seog
authors.initials: HS
authors.lastname: Seo
authors.affiliation: Department of Chemistry, Yonsei University, Seoul, 03722, Republic of Korea. mhmoon@yonsei.ac.kr.
authors.forename: Myeong Hee
authors.initials: MH
authors.lastname: Moon
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis*; D036103:Nanotechnology / Q000379:methods; D065626:Non-alcoholic Fatty Liver Disease / Q000378:metabolism*; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers; D008055:Lipids
keywords: Hepatic lipid; High-cholesterol diet; Inflammation; Nonalcoholic fatty liver disease; nUPLC-ESI-MS/MS