Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Sampling and Analysis of Low Molecular Weight Volatile Metabolites in Cellular Headspace and Mouse Breath

Version 1 : Received: 29 April 2022 / Approved: 6 May 2022 / Online: 6 May 2022 (09:27:04 CEST)

A peer-reviewed article of this Preprint also exists.

Issitt, T.; Sweeney, S.T.; Brackenbury, W.J.; Redeker, K.R. Sampling and Analysis of Low-Molecular-Weight Volatile Metabolites in Cellular Headspace and Mouse Breath. Metabolites 2022, 12, 599. Issitt, T.; Sweeney, S.T.; Brackenbury, W.J.; Redeker, K.R. Sampling and Analysis of Low-Molecular-Weight Volatile Metabolites in Cellular Headspace and Mouse Breath. Metabolites 2022, 12, 599.

Journal reference: Metabolites 2022, 12, 599
DOI: 10.3390/metabo12070599

Abstract

Volatile compounds, abundant in breath, can be used to accurately diagnose and monitor a range of medical conditions. This offers a non-invasive, low-cost approach with screening applications; however, uptake of this diagnostic approach has been limited by conflicting published outcomes. Most published reports rely on large scale screening of the public, at single time points and without reference to ambient air. Here, we present a novel approach to volatile sampling from cellular headspace and mouse breath that incorporates multi-time point analysis and ambient air subtraction revealing compound flux as an effective proxy of active metabolism. This approach to investigating breath volatiles offers a new avenue for disease biomarker discovery and diagnosis. Using gas chromatography mass spectrometry (GC/MS), we focus on low molecular weight, metabolic substrate/by-product compounds and demonstrate that this non-invasive technique is sensitive (reproducible at ~1 µg cellular protein, or ~500,000 cells) and capable of precisely determining cell type, status and treatment. Isolated cellular models represent components of larger mammalian systems and we show that stress- and pathology-indicative compounds are detectable in mice, supporting further investigation using this methodology as a tool to identify volatile targets in human patients.

Keywords

Volatile organic compound; VOC; headspace; breath; breath biomarker; volatile metabolite; breath diagnosis

Subject

MEDICINE & PHARMACOLOGY, General Medical Research

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