Breath Sampling and Patient Considerations for Clinical Implementation: A Comparative Study

This study presents a comprehensive comparison of three exhaled breath sampling methods: ReCIVA, Tedlar bags, and BioVoc. To rigorously evaluate these methods, we employed the Peppermint Initiative, a standardized protocol designed to induce a temporary alteration in the exhaled breath profile. This approach allowed us to track the washout curves of volatile organic compounds (VOCs) following the ingestion of peppermint oil. The collected samples were analyzed using two-dimensional gas chromatography (GC×GC) coupled with high- and medium-resolution mass spectrometry, ensuring detailed and robust analysis. In addition to determining the most effective and reliable sampling technique for clinical use, this study also provides an overview of the commonly used methods, highlighting their strengths and weaknesses in a clinical context as the breathomics community moves towards standardizing collection procedures. This study is currently ongoing. 5 voluntary participants for each sampling method participated to the study. Samples were injected following strict QC protocols to ensure that variability was not instrument-related. Preliminary statistical analysis suggests that variability is induced by the sampling method, potentially linked to adsorption processes, although similar washout curves across all participants support the Peppermint Initiative as a reference for breath sampling assessments. We also considered patient comfort and gathered feedback from hospital staff involved in sampling processes to ensure that the selected method is both analytically sound and clinically practical. The primary goal of this study is to deliver a detailed evaluation of these methods concerning their potential clinical application.