title: Multiplexed peptide analysis for kinetic measurements of major human apolipoproteins by LC/MS/MS.
abstract: A multiplexed assay was developed by MS to analyze, in a single run, six major human Apos involved in lipoprotein metabolism: ApoA-I, ApoA-II, ApoB100, ApoC-II, ApoC-III, and ApoE. This method was validated in vivo in six subjects who received a 14 h constant infusion of [5,5,5-(2)H3]L-leucine at 10 μM/kg/h. Plasma lipoprotein fractions were isolated from collected blood samples and were digested with trypsin. Proteotypic peptides were subsequently analyzed by LC/MS/MS. Enrichment measurement data were compared with those obtained by the standard method using GC/MS. The required time to obtain the LC/MS/MS data was less than that needed for GC/MS. The enrichments from both methods were correlated for ApoA-I (r = 0.994; P < 0.0001) and ApoB100 (r = 0.999; P < 0.0001), and the Bland-Altman plot confirmed the similarity of the two methods. Intra- and inter-assay variability calculated for the six Apos of interest did not exceed 10.7 and 12.5%, respectively, and kinetic parameters were similar and/or in agreement with previously reported data. Therefore, LC/MS/MS can be considered as a useful tool for human Apo kinetic studies using stable isotopes.
authors.affiliation: INRA, UMR 1280, Physiologie des Adaptations Nutritionnelles, CHU Hôtel-Dieu, F-44000 Nantes, France CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France.
authors.forename: Mikaël
authors.initials: M
authors.lastname: Croyal
authors.affiliation: INRA, UMR 1280, Physiologie des Adaptations Nutritionnelles, CHU Hôtel-Dieu, F-44000 Nantes, France CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France.
authors.forename: Fanta
authors.initials: F
authors.lastname: Fall
authors.affiliation: INRA, UMR 1280, Physiologie des Adaptations Nutritionnelles, CHU Hôtel-Dieu, F-44000 Nantes, France CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Ferchaud-Roucher
authors.affiliation: CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France.
authors.forename: Maud
authors.initials: M
authors.lastname: Chétiveaux
authors.affiliation: CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France.
authors.forename: Yassine
authors.initials: Y
authors.lastname: Zaïr
authors.affiliation: INRA, UMR 1280, Physiologie des Adaptations Nutritionnelles, CHU Hôtel-Dieu, F-44000 Nantes, France CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France.
authors.forename: Khadija
authors.initials: K
authors.lastname: Ouguerram
authors.affiliation: INRA, UMR 1280, Physiologie des Adaptations Nutritionnelles, CHU Hôtel-Dieu, F-44000 Nantes, France CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France Department of Endocrinology, Metabolic Diseases, and Nutrition, G and R Laennec Hospital, F-44093 Nantes, France michel.krempf@univ-nantes.fr.
authors.forename: Michel
authors.initials: M
authors.lastname: Krempf
authors.affiliation: INRA, UMR 1280, Physiologie des Adaptations Nutritionnelles, CHU Hôtel-Dieu, F-44000 Nantes, France CRNHO, West Human Nutrition Research Center, CHU, F-44093 Nantes, France Department of Endocrinology, Metabolic Diseases, and Nutrition, G and R Laennec Hospital, F-44093 Nantes, France.
authors.forename: Estelle
authors.initials: E
authors.lastname: Nobécourt
mesh_terms: D001053:Apolipoproteins / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D008297:Male; D008875:Middle Aged; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001053:Apolipoproteins; D010446:Peptide Fragments
keywords: lipoproteins/kinetics; lipoproteins/metabolism; liquid chromatography-tandem mass spectrometry; multiplexed assay; nutrition protein; stable isotope