The peppermint breath test: a benchmarking protocol for breath sampling and analysis using GC-MS

Maxim Wilkinson, Iain R White, Katie Hamshere, Olaf Holz, Sven Schuchardt, Francesca G Bellagambi, Tommaso Lomonaco, Denise Biagini, Fabio Di Francesco, Stephen J Fowler

Research output: Contribution to journalArticlepeer-review

Abstract

Exhaled breath contains hundreds of volatile organic compounds (VOCs) which offers the potential for diagnosing and monitoring a wide range of diseases. As the breath research field has grown, sampling and analytical practices have become highly varied between groups. Standardisation would allow meta-analyses of data from multiple studies and greater confidence in published results. The Peppermint Consortium has been formed to address this task of standardisation. In the current study we aimed to generate initial benchmark values for thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) analysis of breath samples containing peppermint-derived VOCs. Headspace analysis of peppermint oil capsules was performed to determine compounds of interest. Ten healthy participants were recruited by three groups. Each participant provided a baseline breath sample prior to taking a peppermint capsule, with further samples collected at 60, 90, 165, 285 and 360 min following ingestion. Sampling and analytical protocols were different for each institution, in line with their usual practice. Samples were analysed by TD-GC-MS and benchmarking values determined for the time taken for detected peppermint VOCs to return to baseline values. Sixteen compounds were identified in the capsule headspace. Additionally, 2,3-dehydro-1,8-cineole was uniquely found in the breath samples, with a washout profile that suggested it was a product of peppermint metabolism. Five compounds (α-pinene, β-pinene, eucalyptol, menthol and menthone) were quantified by all three groups. Differences in recovery were observed between the groups, particularly for menthone and menthol. The average time taken for VOCs to return to baseline was selected as the benchmark and were 441, 648, 1736, 643 and 375 min for α-pinene, β-pinene, eucalyptol, menthone and menthol respectively. An initial set of easy-to-measure benchmarking values for assessing the performance of TD-GC-MS systems for the analysis of VOCs in breath is presented. These values will be updated when more groups provide additional data.

Original languageEnglish
JournalJournal of Breath Research
DOIs
Publication statusPublished - 22 Jan 2021

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