Extending PTR based breath analysis to real-time monitoring of reactive volatile organic compounds.

Reactive exhaled volatile organic compounds (VOCs) such as nitrogen- and sulfur-containing substances may be related to diseases, metabolic processes and bacterial activity. As these compounds may interact with any surface of the analytical system, time-resolved monitoring and reliable quantification is difficult. We describe a proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS) based analytical method for direct breath-resolved monitoring of reactive compounds. Aliphatic amines were used as test substances. Matrix adapted gas standards were generated by means of a liquid calibration unit. Calibration conditions were adapted in terms of materials, temperature and equilibration time. PTR-ToF-MS conditions were optimized in terms of inlet materials, transfer line and drift tube temperature and drift tube reduced electric field (E/N). Optimized PTR conditions in combination with inert materials and high temperatures considerably reduced the interactions of compounds with the surfaces of the analytical system. Good linearity (R2 > 0.99, RSDs < 5%) with LODs between 0.15 ppbV and 1.23 ppbV and LOQs between 0.24 ppbV and 1.94 ppbV could be achieved. The method was then applied to breath-resolved monitoring of reactive compounds in 17 healthy subjects after high and low oral protein challenge. Exhaled concentrations of trimethylamine, indole, methanethiol, dimethylsulfide, acetone, 2-propanol, 2-butanone and phenol showed significant changes after protein intake. Methanethiol concentrations increased 6-fold within minutes after the protein intake. Optimization of methods and instrument design enabled reliable breath-resolved PTR-MS based analysis of exhaled reactive VOCs in the sub-ppbV range. Continuous in vivo monitoring of exhaled amines and sulphur containing compounds may provide novel non-invasive insight into endogenous and gut bacteria driven protein metabolism.

Continuous real time breath gas monitoring in the clinical environment by proton-transfer-reaction-time-of-flight-mass spectrometry.

Analysis of volatile organic compounds (VOCs) in breath holds great promise for noninvasive diagnostic applications. However, concentrations of VOCs in breath may change quickly, and actual and previous uptakes of exogenous substances, especially in the clinical environment, represent crucial issues. We therefore adapted proton-transfer-reaction-time-of-flight-mass spectrometry for real time breath analysis in the clinical environment. For reasons of medical safety, a 6 m long heated silcosteel transfer line connected to a sterile mouth piece was used for breath sampling from spontaneously breathing volunteers and mechanically ventilated patients. A time resolution of 200 ms was applied. Breath from mechanically ventilated patients was analyzed immediately after cardiac surgery. Breath from 32 members of staff was analyzed in the post anesthetic care unit (PACU). In parallel, room air was measured continuously over 7 days. Detection limits for breath-resolved real time measurements were in the high pptV/low ppbV range. Assignment of signals to alveolar or inspiratory phases was done automatically by a matlab-based algorithm. Quickly and abruptly occurring changes of patients' clinical status could be monitored in terms of breath-to-breath variations of VOC (e.g. isoprene) concentrations. In the PACU, room air concentrations mirrored occupancy. Exhaled concentrations of sevoflurane strongly depended on background concentrations in all participants. In combination with an optimized inlet system, the high time and mass resolution of PTR-ToF-MS provides optimal conditions to trace quick changes of breath VOC profiles and to assess effects from the clinical environment.

Non-Invasive O-Toluidine Monitoring during Regional Anaesthesia with Prilocaine and Detection of Accidental Intravenous Injection in an Animal Model.

Regional anaesthesia is well established as a standard method in clinical practice. Currently, the local anaesthetics of amino-amide types such as prilocaine are frequently used. Despite routine use, complications due to overdose or accidental intravenous injection can arise. A non-invasive method that can indicate such complications early would be desirable. Breath gas analysis offers great potential for the non-invasive monitoring of drugs and their volatile metabolites. The physicochemical properties of o-toluidine, the main metabolite of prilocaine, allow its detection in breath gas. Within this study, we investigated whether o-toluidine can be monitored in exhaled breath during regional anaesthesia in an animal model, if correlations between o-toluidine and prilocaine blood levels exist and if accidental intravenous injections are detectable by o-toluidine breath monitoring. Continuous o-toluidine monitoring was possible during regional anaesthesia of the cervical plexus and during simulated accidental intravenous injection of prilocaine. The time course of exhaled o-toluidine concentrations considerably differed depending on the injection site. Intravenous injection led to an immediate increase in exhaled o-toluidine concentrations within 2 min, earlier peak and higher maximum concentrations, followed by a faster decay compared to regional anaesthesia. The strength of correlation of blood and breath parameters depended on the injection site. In conclusion, real time monitoring of o-toluidine in breath gas is possible by means of PTR-ToF-MS. Since simulated accidental intravenous injection led to an immediate increase in exhaled o-toluidine concentrations within 2 min and higher maximum concentrations, monitoring exhaled o-toluidine may potentially be applied for the non-invasive real-time detection of accidental intravenous injection of prilocaine.

Effects of elevated oxygen levels on VOC analysis by means of PTR-ToF-MS.

Proton-transfer-reaction-time-of-flight-mass-spectrometry (PTR-ToF-MS) is a powerful tool for real-time monitoring of volatile organic compound (VOC) profiles in human breath. However, varying oxygen concentrations in the sample matrix may influence results from VOC analysis by PTR-ToF-MS. Elevated oxygen concentrations are likely to occur in clinical settings, but also when respiratory masks or breathing apparatus are used (e.g. in scuba diving, aviation, firefighting). Oxygen concentration may vary between subjects or within the course of a measurement or study and thus bias results. We systematically assessed the effect of high O2 concentrations (up to 90%) in the sample matrix on the results of PTR-MS analysis for a pattern of VOCs including aromatics, aldehydes and ketones in dry and humid samples. In vivo experiments in healthy volunteers and mechanically ventilated animals were done to test the effect under real-life conditions. H3O+ count significantly decreased by more than 40% when the amount of oxygen in the sample matrix was increased. Almost all investigated VOCs were significantly effected by varying oxygen concentrations and differences in signal intensities of more than 50% could be observed. The effect was generally more pronounced in dry samples but still significant under humid conditions. A linear dependency of sensitivity on the oxygen concentration in the sample matrix was observed for a number of VOCs (e.g. aldehydes) possibly enabling a factor based correction. VOC intensities were also influenced under in vivo conditions, e.g. ethanol decreased up to 71%. When PTR-MS analysis is carried out under oxygen supply, these issues need to be carefully considered.

VOC breath profile in spontaneously breathing awake swine during Influenza A infection.

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.

Monitoring of breath VOCs and electrical impedance tomography under pulmonary recruitment in mechanically ventilated patients.

Analysis of exhaled VOCs can provide information on physiology, metabolic processes, oxidative stress and lung diseases. In critically ill patients, VOC analysis may be used to gain complimentary information beyond global clinical parameters. This seems especially attractive in mechanically ventilated patients frequently suffering from impairment of gas exchange. This study was intended to assess (a) the effects of recruitment maneuvers onto VOC profiles, (b) the correlations between electrical impedance tomography (EIT) data and VOC profiles and (c) the effects of recruitment onto distribution of ventilation. Eleven mechanically ventilated patients were investigated during lung recruitment after cardiac surgery. Continuous breath gas analysis by means of PTR-ToF-MS, EIT and blood gas analyses were performed simultaneously. More than 300 mass traces could be detected and monitored continuously by means of PTR-ToF-MS in every patient. Exhaled VOC concentrations varied with recruitment induced changes in minute ventilation and cardiac output. Ammonia exhalation depended on blood pH. The improvement in dorsal lung ventilation during recruitment ranged from 9% to 110%. Correlations between exhaled concentrations of acetone, isoprene, benzene sevoflurane and improvement in regional ventilation during recruitment were observed. Extent and quality of these correlations depended on physico-chemical properties of the VOCs. Combination of continuous real-time breath analysis and EIT revealed correlations between exhaled VOC concentrations and distribution of ventilation. This setup enabled immediate recognition of physiological and therapeutic effects in ICU patients. In a perspective, VOC analysis could be used for non-invasive control and optimization of ventilation strategies.

In Vivo Testing of Extracorporeal Membrane Ventilators: iLA-Activve Versus Prototype I-Lung.

A side-by-side comparison of the decarboxylation efficacy of two pump-driven venovenous extracorporeal lung assist devices, i.e., a first prototype of the new miniaturized ambulatory extracorporeal membrane ventilator, I-lung versus the commercial system iLA-activve for more than a period of 72 hours in a large animal model. Fifteen German Landrace pigs were anesthetized and underwent mechanical hypoventilation to induce severe hypercapnia. Decarboxylation was accomplished by either the I-lung or the iLA-activve via a double lumen catheter in the jugular vein. Sham-operated pigs were not connected to extracorporeal devices. Cardiovascular, respiratory, and metabolic parameters were continuously monitored, combined with periodic arterial blood sampling for subsequent clinical blood diagnostics, such as gas exchange, hemolysis, coagulation parameters, and cytokine profiles. At the termination of the studies, lung tissue was harvested and examined histologically for pulmonary morphology and leukocyte tissue infiltration. Both extracorporeal devices showed high and comparable efficacy with respect to carbon dioxide elimination for more than 72 hours and were not associated with either bleeding events or clotting disorders. Pigs of both groups showed cardiovascular and hemodynamic stability without marked differences to sham-operated animals. Groups also did not differ in terms of inflammatory and metabolic parameters. We established a preclinical in vivo porcine model for comparative long-term testing of I-lung and iLA-activve. The I-lung prototype proved to be safe and feasible, providing adequate decarboxylation without any adverse events. Once translated into the clinical treatment, the new miniaturized and transportable I-lung device might represent a promising tool for treating awake and mobilized patients with decompensated pulmonary disorders.