title: Top-down and bottom-up lipidomic analysis of rabbit lipoproteins under different metabolic conditions using flow field-flow fractionation, nanoflow liquid chromatography and mass spectrometry.
abstract: This study demonstrated the performances of top-down and bottom-up approaches in lipidomic analysis of lipoproteins from rabbits raised under different metabolic conditions: healthy controls, carrageenan-induced inflammation, dehydration, high cholesterol (HC) diet, and highest cholesterol diet with inflammation (HCI). In the bottom-up approach, the high density lipoproteins (HDL) and the low density lipoproteins (LDL) were size-sorted and collected on a semi-preparative scale using a multiplexed hollow fiber flow field-flow fractionation (MxHF5), followed by nanoflow liquid chromatography-ESI-MS/MS (nLC-ESI-MS/MS) analysis of the lipids extracted from each lipoprotein fraction. In the top-down method, size-fractionated lipoproteins were directly infused to MS for quantitative analysis of targeted lipids using chip-type asymmetrical flow field-flow fractionation-electrospray ionization-tandem mass spectrometry (cAF4-ESI-MS/MS) in selected reaction monitoring (SRM) mode. The comprehensive bottom-up analysis yielded 122 and 104 lipids from HDL and LDL, respectively. Rabbits within the HC and HCI groups had lipid patterns that contrasted most substantially from those of controls, suggesting that HC diet significantly alters the lipid composition of lipoproteins. Among the identified lipids, 20 lipid species that exhibited large differences (>10-fold) were selected as targets for the top-down quantitative analysis in order to compare the results with those from the bottom-up method. Statistical comparison of the results from the two methods revealed that the results were not significantly different for most of the selected species, except for those species with only small differences in concentration between groups. The current study demonstrated that top-down lipid analysis using cAF4-ESI-MS/MS is a powerful high-speed analytical platform for targeted lipidomic analysis that does not require the extraction of lipids from blood samples.
authors.affiliation: Department of Chemistry, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Seul Kee
authors.initials: SK
authors.lastname: Byeon
authors.affiliation: Department of Chemistry, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Jin Yong
authors.initials: JY
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Ju Yong
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Future Convergence Research Division, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea.
authors.forename: Bong Chul
authors.initials: BC
authors.lastname: Chung
authors.affiliation: Cardiovascular Center, Korea University Guro Hospital, Seoul 152-703, Republic of Korea.
authors.forename: Hong Seog
authors.initials: HS
authors.lastname: Seo
authors.affiliation: Department of Chemistry, Yonsei University, Seoul 120-749, Republic of Korea. Electronic address: mhmoon@yonsei.ac.kr.
authors.forename: Myeong Hee
authors.initials: MH
authors.lastname: Moon
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D031521:Fractionation, Field Flow / Q000379:methods; D008074:Lipoproteins / Q000097:blood*; D008075:Lipoproteins, HDL / Q000097:blood; D008077:Lipoproteins, LDL / Q000097:blood; D011817:Rabbits; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL
keywords: Carrageenan; High cholesterol diet; Lipoproteins; Rabbit serum; Top-down lipidomics; nLC-ESI-MS/MS