%0 Journal Article
%T Combination of real-time and hyphenated mass spectrometry for improved characterisation of exhaled breath biomarkers in clinical research.
%A Lamy E
%A Roquencourt C
%A Zhou B
%A Salvator H
%A Moine P
%A Annane D
%A Devillier P
%A Bardin E
%A Grassin-Delyle S
%J Anal Bioanal Chem
%V 416
%N 22
%D 2024 Sep 9
%M 38980330
%F 4.478
%R 10.1007/s00216-024-05421-7
%X Exhaled breath volatilomics is a powerful non-invasive tool for biomarker discovery in medical applications, but compound annotation is essential for pathophysiological insights and technology transfer. This study was aimed at investigating the interest of a hybrid approach combining real-time proton transfer reaction-time-of-flight mass spectrometry (PTR-TOF-MS) with comprehensive thermal desorption-two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (TD-GCxGC-TOF-MS) to enhance the analysis and characterization of VOCs in clinical research, using COVID-19 as a use case. VOC biomarker candidates were selected from clinical research using PTR-TOF-MS fingerprinting in patients with COVID-19 and matched to the Human Breathomic Database. Corresponding analytical standards were analysed using both a liquid calibration unit coupled to PTR-TOF-MS and TD-GCxGC-TOF-MS, together with confirmation on new clinical samples with TD-GCxGC-TOF-MS. From 26 potential VOC biomarkers, 23 were successfully detected with PTR-TOF-MS. All VOCs were successfully detected using TD-GCxGC-TOF-MS, providing effective separation of highly chemically related compounds, including isomers, and enabling high-confidence annotation based on two-dimensional chromatographic separation and mass spectra. Four VOCs were identified with a level 1 annotation in the clinical samples. For future applications, the combination of real-time PTR-TOF-MS and comprehensive TD-GCxGC-TOF-MS, at least on a subset of samples from a whole study, would enhance the performance of VOC annotation, offering potential advancements in biomarker discovery for clinical research.