Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 173, Issue -, Pages 1-17Publisher
ELSEVIER
DOI: 10.1016/j.jpba.2019.05.002
Keywords
Bile acids; Profiling; Postprandial kinetics; Metabotypes; Triple Quad; Orbitrap
Categories
Funding
- Project Cabala_diethealth
- European Union's Horizon2020 research and innovation [696295]
- Autonomous Province of Trento, ADP 2018
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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 mu A biofluid-aliquots, followed by a properly optimised 27 min-chromatographic run. The method provided high sensitivity (limits of detection 0.0002-0.03 mu M, limits of quantitation 0.0007-0.11 mu M), linearity (R-2>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. (C) 2019 The Authors. Published by Elsevier B.V.
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