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Isotopic dilution method for bile acid profiling reveals new sulfate glycine-conjugated dihydroxy bile acids and glucuronide bile acids in serum.

Authors
  • Ulaszewska, Maria M1
  • Mancini, Andrea2
  • Garcia-Aloy, Mar2
  • Del Bubba, Massimo3
  • Tuohy, Kieran Micheal2
  • Vrhovsek, Urska2
  • 1 Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach (FEM), Via Mach 1, 38010, San Michele all'Adige, Trento, Italy. Electronic address: [email protected] , (Italy)
  • 2 Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach (FEM), Via Mach 1, 38010, San Michele all'Adige, Trento, Italy. , (Italy)
  • 3 Department of Chemistry, University of Florence, Via della Lastruccia 3, 50019, Sesto Fiorentino, Florence, Italy. , (Italy)
Type
Published Article
Journal
Journal of pharmaceutical and biomedical analysis
Publication Date
Sep 05, 2019
Volume
173
Pages
1–17
Identifiers
DOI: 10.1016/j.jpba.2019.05.002
PMID: 31100508
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

An ultrahigh performance liquid chromatography tandem mass spectrometry method (UHPLC-MS/MS) was developed for the determination of 41 target and 8 additional bile acids isomers (BAs) in biological fluids. BAs were analysed by solid-phase extraction on 50 μL biofluid-aliquots, followed by a properly optimised 27 min-chromatographic run. The method provided high sensitivity (limits of detection 0.0002-0.03 μM, limits of quantitation 0.0007-0.11 μM), linearity (R2>0.99) and precision (relative standard deviations ≤16%). A strategy of scheduled/ unscheduled injections of real samples together with neutral loss (80 Da and 176 Da) scans allowed us to find additional bile acid isomers not a priori included in the method, while high resolution full scan and MS/MS fragmentation analysis confirmed their structural adherence to the bile acid family. Moreover this is the first study quantifying four sulfate glycine conjugated-dihydroxy bile acid isomers, independently of the diet and postprandial time. Application to a dietary intervention kinetic study confirmed the existence of possible metabotypes amongst the study population (n = 20). A trend differentiating males from females was observed suggesting that serum samples from women contained smaller amounts of certain bile acids. Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

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