Rapid concentration changes due to physiological or pathophysiological effects rather than appearance of unique disease biomarkers are important for clinical application of breath research. Simple maneuvers such as breath holding may significantly affect breath biomarker concentrations. In this study, exhaled volatile organic compound (VOC) concentrations were assessed in real time before and after different breath holding maneuvers. Continuous breath-resolved measurements (PTR-ToF-MS-8000) were performed in 31 healthy human subjects in a side-stream sampling mode. After 1 min of tidal breathing participants held their breath for 10, 20, 40, 60 s and as long as possible. Afterwards they continued to breathe normally for another minute. VOC profiles could be monitored in real time by assigning online PTR-ToF-MS data to alveolar or inspired phases of breath. Sudden and profound changes of exhaled VOC concentrations were recorded after different breath holding maneuvers. VOC concentrations returned to base line levels 10-20 s after breath holding. Breath holding induced concentration changes depended on physico-chemical properties of the substances. When substance concentrations were normalized onto end-tidal CO2 content, variation of acetone concentrations decreased, whereas variations of isoprene concentrations were not affected. As the effects of breathing patterns on exhaled substance concentrations depend on individual substance properties, sampling procedures have to be validated for each compound by means of appropriate real-time analysis. Normalization of exhaled concentrations onto exhaled CO2 is only valid for substances having similar physico-chemical properties as CO2.

Concentrations of exhaled volatile organic compounds (VOCs) may depend not only on biochemical or pathologic processes but also on physiological parameters. As breath sampling may be done in different body positions, effects of the sampling position on exhaled VOC concentrations were investigated by means of real-time mass spectrometry. Breaths from 15 healthy volunteers were analyzed in real-time by PTR-ToF-MS-8000 during paced breathing (12/min) in a continuous side-stream mode. We applied two series of body positions (setup 1: sitting, standing, supine, and sitting; setup 2: supine, left lateral, right lateral, prone, and supine). Each position was held for 2 min. Breath VOCs were quantified in inspired and alveolar air by means of a custom-made algorithm. Parallel monitoring of hemodynamics and capnometry was performed noninvasively. In setup 1, when compared to the initial sitting position, normalized mean concentrations of isoprene, furan, and acetonitrile decreased by 24%, 26%, and 9%, respectively, during standing and increased by 63%, 36%, and 10% during lying mirroring time profiles of stroke volume and pET-CO2. In contrast, acetone and H2S concentrations remained almost constant. In setup 2, when compared to the initial supine position, mean alveolar concentrations of isoprene and furan increased significantly up to 29% and 16%, respectively, when position was changed from lying on the right side to the prone position. As cardiac output and stroke volume decreased at that time, the reasons for the observed concentrations changes have to be linked to the ventilation/perfusion ratio or compartmental distribution rather than to perfusion alone. During final postures, all VOC concentrations, hemodynamics, and pET-CO2 returned to baseline. Exhaled blood-borne VOC profiles changed due to body postures. Changes depended on cardiac stroke volume, origin, compartmental distribution and physico-chemical properties of the substances. Patients' positions and cardiac output have to be controlled when concentrations of breath VOCs are to be interpreted in terms of biomarkers.

Influenza is one of the most common causes of virus diseases worldwide. Virus detection requires determination of Influenza RNA in the upper respiratory tract. Efficient screening is not possible in this way. Analysis of volatile organic compounds (VOCs) in breath holds promise for non-invasive and fast monitoring of disease progression. Breath VOC profiles of 14 (3 controls and 11 infected animals) swine were repeatedly analyzed during a complete infection cycle of Influenza A under high safety conditions. Breath VOCs were pre-concentrated by means of needle trap micro-extraction and analysed by gas chromatography mass spectrometry before infection, during virus presence in the nasal cavity, and after recovery. Six VOCs could be related to disease progression: acetaldehyde, propanal, n-propyl acetate, methyl methacrylate, styrene and 1,1-dipropoxypropane. As early as on day four after inoculation, when animals were tested positive for Influenza A, differentiation between control and infected animals was possible. VOC based information on virus infection could enable early detection of Influenza A. As VOC analysis is completely non-invasive it has potential for large scale screening purposes. In a perspective, breath analysis may offer a novel tool for Influenza monitoring in human medicine, animal health control or border protection.