Alveolar gradients in breath analysis. A pilot study with comparison of room air and inhaled air by simultaneous measurements using ion mobility spectrometry.

Analyzing exhaled breath samples, especially using a highly sensitive method such as MCC/IMS (multi-capillary column/ion mobility spectrometry), may also detect analytes that are derived from exogenous production. In this regard, there is a discussion about the optimal interpretation of exhaled breath, either by considering volatile organic compounds (VOCs) only in exhaled breath or by additionally considering the composition of room air and calculating the alveolar gradients. However, there are no data on whether the composition and concentration of VOCs in room air are identical to those in truly inhaled air directly before analyzing the exhaled breath. The current study aimed to determine whether the VOCs in room air, which are usually used for the calculation of alveolar gradients, are identical to the VOCs in truly inhaled air. For the measurement of inhaled air and room air, two IMS, each coupled with an MCC that provided a pre-separation of the VOCs, were used in parallel. One device was used for sampling room air and the other for sampling inhaled air. Each device was coupled with a newly invented system that cleaned room air and provided a clean carrier gas, whereas formerly synthetic air had to be used as a carrier gas. In this pilot study, a healthy volunteer underwent three subsequent runs of sampling of inhaled air and simultaneous sampling and analysis of room air. Three of the selected 11 peaks (P4-unknown, P5-1-Butanol, and P9-Furan, 2-methyl-) had significantly higher intensities during inspiration than in room air, and four peaks (P1-1-Propanamine, N-(phenylmethylene), P2-2-Nonanone, P3-Benzene, 1,2,4-trimethyl-, and P11-Acetyl valeryl) had higher intensities in room air. Furthermore, four peaks (P6-Benzaldehyde, P7-Pentane, 2-methyl-, P8-Acetone, and P10-2-Propanamine) showed inconsistent differences in peak intensities between inhaled air and room air. To the best of our knowledge, this is the first study to compare simultaneous sampling of room air and inhaled air using MCC/IMS. The simultaneous measurement of inhaled air and room air showed that using room air for the calculation of alveolar gradients in breath analysis resulted in different alveolar gradient values than those obtained by measuring truly inhaled air.

A proof of concept study for the differentiation of SARS-CoV-2, hCoV-NL63, and IAV-H1N1 in vitro cultures using ion mobility spectrometry.

Rapid, high-throughput diagnostic tests are essential to decelerate the spread of the novel coronavirus disease 2019 (COVID-19) pandemic. While RT-PCR tests performed in centralized laboratories remain the gold standard, rapid point-of-care antigen tests might provide faster results. However, they are associated with markedly reduced sensitivity. Bedside breath gas analysis of volatile organic compounds detected by ion mobility spectrometry (IMS) may enable a quick and sensitive point-of-care testing alternative. In this proof-of-concept study, we investigated whether gas analysis by IMS can discriminate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from other respiratory viruses in an experimental set-up. Repeated gas analyses of air samples collected from the headspace of virus-infected in vitro cultures were performed for 5 days. A three-step decision tree using the intensities of four spectrometry peaks correlating to unidentified volatile organic compounds allowed the correct classification of SARS-CoV-2, human coronavirus-NL63, and influenza A virus H1N1 without misassignment when the calculation was performed with data 3 days post infection. The forward selection assignment model allowed the identification of SARS-CoV-2 with high sensitivity and specificity, with only one of 231 measurements (0.43%) being misclassified. Thus, volatile organic compound analysis by IMS allows highly accurate differentiation of SARS-CoV-2 from other respiratory viruses in an experimental set-up, supporting further research and evaluation in clinical studies.

Statistical analysis of MCC-IMS data for two group comparisons-an exemplary study on two devices.

The Multi-capillary-column-Ion-mobility-spectrometry (MCC-IMS) technology for measuring breath gas can be used for distinguishing between healthy and diseased subjects or between different types of diseases. The statistical methods for classifying the corresponding breath samples typically neglects potential confounding clinical and technical variables, reducing both accuracy and generalizability of the results. Especially measuring samples on different technical devices can heavily influence the results. We conducted a controlled breath gas study including 49 healthy volunteers to evaluate the effect of the variables sex, smoking habits and technical device. Every person was measured twice, once before and once after consuming a glass of orange juice. The two measurements were obtained on two different devices. The evaluation of the MCC-IMS data regarding metabolite detection was performed once using the software VisualNow, which requires manual interaction, and once using the fully automated algorithm SGLTR-DBSCAN. We present statistical solutions, peak alignment and scaling, to adjust for the different devices. For the other potential confounders sex and smoking, in our study no significant influence was identified.

Adherence of volatile propofol to various types of plastic tubing.

Propofol is an intravenous anesthetic. Currently, it is not possible to routinely measure blood concentration of the drug in real time. However, multi-capillary column ion-mobility spectrometry of exhaled gas can estimate blood propofol concentration. Unfortunately, adhesion of volatile propofol on plastic materials complicates measurements. Therefore, it is necessary to consider the extent to which volatile propofol adheres to various plastics used in sampling tubing. Perfluoralkoxy (PFA), polytetrafluorethylene (PTFE), polyurethane (PUR), silicone, and Tygon tubing were investigated in an experimental setting using a calibration gas generator (HovaCAL). Propofol gas was measured for one hour at 26 °C, 50 °C, and 90 °C tubing temperature. Test tubing segments were then flushed with N2 to quantify desorption. PUR and Tygon sample tubing absorbed all volatile propofol. The silicone tubing reached the maximum propofol concentration after 119 min which was 29 min after propofol gas exposure stopped. The use of PFA or PTFE tubing produced comparable and reasonably accurate propofol measurements. The desaturation time for the PFA was 10 min shorter at 26 °C than for PTFE. PFA tubing thus seems most suitable for measurement of volatile propofol, with PTFE as an alternative.

Influence of the respirator on volatile organic compounds: an animal study in rats over 24 hours.

Long-term animal studies are needed to accomplish measurements of volatile organic compounds (VOCs) for medical diagnostics. In order to analyze the time course of VOCs, it is necessary to ventilate these animals. Therefore, a total of 10 male Sprague-Dawley rats were anaesthetized and ventilated with synthetic air via tracheotomy for 24 h. An ion mobility spectrometry coupled to multi-capillary columns (MCC-IMS) was used to analyze the expired air. To identify background contaminations produced by the respirator itself, six comparative measurements were conducted with ventilators only. Overall, a number of 37 peaks could be detected within the positive mode. According to the ratio peak intensity rat/ peak intensity ventilator blank, 22 peaks with a ratio >1.5 were defined as expired VOCs, 12 peaks with a ratio between 0.5 and 1.5 as unaffected VOCs, and three peaks with a ratio <0.5 as resorbed VOCs. The peak intensity of 12 expired VOCs changed significantly during the 24 h measurement. These results represent the basis for future intervention studies. Notably, online VOC analysis with MCC-IMS is possible over 24 h in ventilated rats and allows different experimental approaches.

Review on ion mobility spectrometry. Part 1: current instrumentation.

Ion Mobility Spectrometry (IMS) is a widely used and 'well-known' technique of ion separation in the gaseous phase based on the differences in ion mobilities under an electric field. All IMS instruments operate with an electric field that provides space separation, but some IMS instruments also operate with a drift gas flow that provides also a temporal separation. In this review we will summarize the current IMS instrumentation. IMS techniques have received an increased interest as new instrumentation and have become available to be coupled with mass spectrometry (MS). For each of the eight types of IMS instruments reviewed it is mentioned whether they can be hyphenated with MS and whether they are commercially available. Finally, out of the described devices, the six most-consolidated ones are compared. The current review article is followed by a companion review article which details the IMS hyphenated techniques (mainly gas chromatography and mass spectrometry) and the factors that make the data from an IMS device change as a function of device parameters and sampling conditions. These reviews will provide the reader with an insightful view of the main characteristics and aspects of the IMS technique.

Review on ion mobility spectrometry. Part 2: hyphenated methods and effects of experimental parameters.

Ion Mobility Spectrometry (IMS) is a widely used and 'well-known' technique of ion separation in the gaseous phase based on the differences of ion mobilities under an electric field. This technique has received increased interest over the last several decades as evidenced by the pace and advances of new IMS devices available. In this review we explore the hyphenated techniques that are used with IMS, specifically mass spectrometry as an identification approach and a multi-capillary column as a pre-separation approach. Also, we will pay special attention to the key figures of merit of the ion mobility spectrum and how data sets are treated, and the influences of the experimental parameters on both conventional drift time IMS (DTIMS) and miniaturized IMS also known as high Field Asymmetric IMS (FAIMS) in the planar configuration. The present review article is preceded by a companion review article which details the current instrumentation and contains the sections that configure both conventional DTIMS and FAIMS devices. These reviews will give the reader an insightful view of the main characteristics and aspects of the IMS technique.

Two different approaches for pharmacokinetic modeling of exhaled drug concentrations.

Online measurement of drug concentrations in patient's breath is a promising approach for individualized dosage. A direct transfer from breath- to blood-concentrations is not possible. Measured exhaled concentrations are following the blood-concentration with a delay in non-steady-state situations. Therefore, it is necessary to integrate the breath-concentration into a pharmacological model. Two different approaches for pharmacokinetic modelling are presented. Usually a 3-compartment model is used for pharmacokinetic calculations of blood concentrations. This 3-compartment model is extended with a 2-compartment model based on the first compartment of the 3-compartment model and a new lung compartment. The second approach is to calculate a time delay of changes in the concentration of the first compartment to describe the lung-concentration. Exemplarily both approaches are used for modelling of exhaled propofol. Based on time series of exhaled propofol measurements using an ion-mobility-spectrometer every minute for 346 min a correlation of calculated plasma and the breath concentration was used for modelling to deliver R(2) = 0.99 interdependencies. Including the time delay modelling approach the new compartment coefficient k(e0lung) was calculated to k(e0lung) = 0.27 min(-1) with R(2) = 0.96. The described models are not limited to propofol. They could be used for any kind of drugs, which are measurable in patient's breath.

Wash-out of ambient air contaminations for breath measurements.

In breath analysis, ambient air contaminations are ubiquitous and difficult to eliminate. This study was designed to investigate the reduction of ambient air background by a lung wash-out with synthetic air. The reduction of the initial ambient air volatile organic compound (VOC) intensity was investigated in the breath of 20 volunteers inhaling synthetic air via a sealed full face mask in comparison to inhaling ambient air. Over a period of 30 minutes, breath analysis was conducted using ion mobility spectrometry coupled to a multi-capillary column. A total of 68 VOCs were identified for inhaling ambient air or inhaling synthetic air. By treatment with synthetic air, 39 VOCs decreased in intensity, whereas 29 increased in comparison to inhaling ambient air. In total, seven VOCs were significantly reduced (P-value < 0.05). A complete wash-out of VOCs in this setting was not observed, whereby a statistically significant reduction up to 65% as for terpinolene was achieved. Our setting successfully demonstrated a reduction of ambient air contaminations from the airways by a lung wash-out with synthetic air.

Multi-capillary column-ion mobility spectrometer (MCC-IMS) breath analysis in ventilated rats: a model with the feasibility of long-term measurements.

Rats are commonly used in medical research as they enable a high grade of standardization. The exhalome of ventilated rats has not as yet been investigated using an ion mobility spectrometer coupled with a multi-capillary column (MCC-IMS). As a first step, a rat model has to be established to measure potential biomarkers in the exhale with long-term settings, allowing constant and continuous analysis of exhaled air in time series. Therefore, eight animals were anaesthetized, prepared and ventilated for 1 h. A total of 73 peaks were directly detected with the IMS chromatogram. Thirty five of them were assigned to the ventilator system and 38 to the animals. Peak intensity varied within three measurements. The intensity of analytes of individual rats varied by a factor of up to 18. This new model will also enable continuous measurements of volatile organic compounds (VOCs) from rat's breath in long-term experiments. It is hoped that, in the future, variability and progression of VOCs can be monitored in different models of diseases using this set-up.

Integrated statistical learning of metabolic ion mobility spectrometry profiles for pulmonary disease identification.

Exhaled air carries information on human health status. Ion mobility spectrometers combined with a multi-capillary column (MCC/IMS) is a well-known technology for detecting volatile organic compounds (VOCs) within human breath. This technique is relatively inexpensive, robust and easy to use in every day practice. However, the potential of this methodology depends on successful application of computational approaches for finding relevant VOCs and classification of patients into disease-specific profile groups based on the detected VOCs. We developed an integrated state-of-the-art system using sophisticated statistical learning techniques for VOC-based feature selection and supervised classification into patient groups. We analyzed breath data from 84 volunteers, each of them either suffering from chronic obstructive pulmonary disease (COPD), or both COPD and bronchial carcinoma (COPD + BC), as well as from 35 healthy volunteers, comprising a control group (CG). We standardized and integrated several statistical learning methods to provide a broad overview of their potential for distinguishing the patient groups. We found that there is strong potential for separating MCC/IMS chromatograms of healthy controls and COPD patients (best accuracy COPD vs CG: 94%). However, further examination of the impact of bronchial carcinoma on COPD/no-COPD classification performance is necessary (best accuracy CG vs COPD vs COPD + BC: 79%). We also extracted 20 high-scoring VOCs that allowed differentiating COPD patients from healthy controls. We conclude that these statistical learning methods have a generally high accuracy when applied to well-structured, medical MCC/IMS data.

[Smelling diseases? A short review on electronic noses]. / Krankheiten erriechen? Eine kurze Übersicht über elektronische Nasen.

Non-invasive pulmonary diagnostics is a promising and interesting field in respiratory medicine. Beside exhaled breath condensate, there is an increasing interest in alternative and faster techniques such as electronic noses (EN). EN aim to mimic or improve the sense of smelling. Different types of EN have been employed in research so far. In addition to ion mobility spectrometry and mass spectrometry, ENs that consist of various biopolymer sensors for the sensing of volatile organic compounds (VOCs) have been tested. VOCs bind to the sensors depending on size, structure, hydrogen binding and polarity. This leads to physical alterations, e. g., swelling resulting in a change of resistance. The smell print represents composite patterns in contrast to single compounds, and the distinction between different categories is achieved by pattern recognition algorithms. Other types of EN like mass spectrometry and ion mobility spectrometry are capable of identifying even single analyte fractions provided that their characteristics have been saved in data repositories. The non-invasive nature, onsite availability and relatively cheap sampling are advantages of ENs that underly the increasing interest in their use for medical purposes. Some promising results have already been published. This review aims to describe the state of the art in brief form.

Ion mobility spectrometry for the detection of volatile organic compounds in exhaled breath of patients with lung cancer: results of a pilot study.

BACKGROUND: Analysis of exhaled breath, especially of volatile organic compounds (VOCs), is of increasing interest in the diagnosis of lung cancer. Compared with other methods of breath analysis, ion mobility spectrometry (IMS) offers a tenfold higher detection rate of VOCs. By coupling the ion mobility spectrometer with a multicapillary column as a pre-separation unit, IMS offers the advantage of an immediate twofold separation of VOCs with visualisation in a three-dimensional chromatogram. The total analysis time is about 500 s compared with gas chromatography/mass spectrometry (GC/MS) of about 1 h. It therefore seemed reasonable to test IMS in breath analysis. METHODS: In a pilot study, 32 patients with lung cancer were subjected to a breath analysis by IMS. Their IMS chromatograms were compared with those of 54 healthy controls. An IMS that was built for special clinical application was used to identify characteristic peaks of VOCs which might be relevant for the diagnosis of lung cancer in exhaled air of 10 ml volume. RESULTS: By a combination of 23 peak regions within the IMS chromatogram, patients with lung cancer, including a patient with carcinoma in situ, were classified and differentiated from healthy persons with an error rate of zero. CONCLUSION: Breath analysis by IMS can detect a discriminating combination of VOCs in patients with lung cancer. By pattern recognition without the need for chemical analysis of the underlying VOCs, IMS has the potential to facilitate lung cancer diagnosis.

MCC/IMS signals in human breath related to sarcoidosis-results of a feasibility study using an automated peak finding procedure.

A feasibility study using an ion mobility spectrometer coupled with a multi-capillary column (MCC) was started to identify characteristic peaks of volatile compounds in exhaled human breath samples of 10 ml volume. The breath of 20 patients with sarcoidosis and suspicion of sarcoidosis because of mediastinal lymph node enlargement was investigated. Using a set of procedures for data processing and scoring a sector of interest was determined within the IMS-chromatogram. It could be shown that a procedure related to a single peak in the IMS-chromatogram delivers differentiation into the two groups of patients with confirmed sarcoidosis and those suffering no sarcoidosis. The potential biomarker is characterized by the following parameters: inverse mobility (1/K(0)) 0.53 ± 0.01 V s cm(-2)-retention time 22 ± 5 s. These results are a first step in breath analysis by MCC/IMS in patients with sarcoidosis by an automated procedure applied to IMS-chromatograms directly.

Ion mobility spectrometry of gas-phase ions from laser ablation of solids in air at ambient pressure.

A mobility spectrometer was used to characterize gas-phase ions produced from laser ablation of solids in air at 100 degrees C and at ambient pressure with a beam focused to a diameter of

Role of undecan-2-one on ethanol-induced apoptosis in HepG2 cells.

Based on the reduced expression of ethanol-oxidizing enzymes in human hepatocellular carcinoma (HepG2) cells, we analyzed the role of nonoxidative metabolites in ethanol-induced apoptosis in HepG2 cells. For this purpose, an analysis of volatile metabolites of ethanol using ion-mobility spectrometry and gas chromatography-mass spectrometry was performed. HepG2 cells exposed to 1 mmol/L ethanol exhibited significant synthesis of undecan-2-one compared to untreated cells. Undecan-2-one is a fatty acid ethyl ester metabolite synthesized through a nonoxidative pathway. Undecan-2-one had a dose-dependent cytotoxic effect on HepG2 cells as shown by release of lactate dehydrogenase (LDH). The most notable finding of this study was the potentiation of ethanol-induced apoptosis demonstrated by an increased apoptotic rate induced by undecan-2-one in ethanol-treated HepG2 cells. The data presented in this study contribute to the better understanding of the molecular mechanisms of ethanol exposure at low concentration in HepG2 cells, a human hepatocellular carcinoma-derived cell line.

Detection of sulfur-free odorants in natural gas using ion mobility spectrometry.

Beside the primary motivation of the public gas suppliers for odorizing natural gas with a sulfur-free odorant, which relates to the image of the environment-friendly fuel, natural gas, competing with low-sulfur heating fuel and diesel, a question of crucial importance of how to detect such sulfur-free odorants comes up. Concerning the replacement of sulfur-containing by sulfur-free odorization, the availability of a fast and sensitive detection method that can, further, be used on-site plays a key role. The minimum concentration of the new sulfur-free odorant Gasodor S-Free (S-Free) in natural gas should be added at a level of at least 8.8 mg m(-3) to assure a significant warning smell. Therefore, a dynamic range between 0 and approx. 25 mg m(-3) must be realised in the rather complex matrix of natural gas. By means of a handheld ion mobility spectrometer, the odorant content in natural gas is determined within less than 80 s total analysis time directly at the gas pipe. The concentration of S-Free is monitored between 4 and 23 mg m(-3) respecting the quality of the natural gas (high- and low-caloric gas). Results of the validation using a gas chromatograph as a reference standard will be discussed in detail.

Ion mobility spectrometry in the diagnosis of sarcoidosis: results of a feasibility study.

A feasibility study with ion mobility spectrometry (IMS) was started to find characteristic peaks of volatile organic compounds in exhaled air of 10 mL sampling volume, which might be relevant for the diagnosis of sarcoidosis. Therefore, breath samples of 9 patients with sarcoidosis and suspicion of sarcoidosis because of mediastinal lymph node enlargement were investigated. The 5 patients with confirmed sarcoidosis showed a highly congruent distribution of metabolites in exhaled air which was different in main component analyses from patients with unspecific mediastinal lymph node enlargement. These results are a first step in breath analysis by IMS in patients with sarcoidosis. The IMS as a new method in breath analysis and the first results of the investigations are presented and discussed in detail.

Ion mobility spectrometry for food quality and safety.

Ion mobility spectrometry is known to be a fast and sensitive technique for the detection of trace substances, and it is increasingly in demand not only for protection against explosives and chemical warfare agents, but also for new applications in medical diagnosis or process control. Generally, a gas phase sample is ionized by help of ultraviolet light, ss-radiation or partial discharges. The ions move in a weak electrical field towards a detector. During their drift they collide with a drift gas flowing in the opposite direction and, therefore, are slowed down depending on their size, shape and charge. As a result, different ions reach the detector at different drift times, which are characteristic for the ions considered. The number of ions reaching the detector are a measure of the concentration of the analyte. The method enables the identification and quantification of analytes with high sensitivity (ng l(-1) range). The selectivity can even be increased - as necessary for the analyses of complex mixtures - using pre-separation techniques such as gas chromatography or multi-capillary columns. No pre-concentration of the sample is necessary. Those characteristics of the method are preserved even in air with up to a 100% relative humidity rate. The suitability of the method for application in the field of food quality and safety - including storage, process and quality control as well as the characterization of food stuffs - was investigated in recent years for a number of representative examples, which are summarized in the following, including new studies as well: (1) the detection of metabolites from bacteria for the identification and control of their growth; (2) process control in food production - beer fermentation being an example; (3) the detection of the metabolites of mould for process control during cheese production, for quality control of raw materials or for the control of storage conditions; (4) the quality control of packaging materials during the production of polymeric materials; and (5) the characterization of products - wine being an example. The challenges of such applications were operation in humid air, fast on-line analyses of complex mixtures, high sensitivity - detection limits have to be, for example, in the range of the odour limits - and, in some cases, the necessity of mobile instrumentation. It can be shown that ion mobility spectrometry is optimally capable of fulfilling those challenges for many applications.

Process analysis using ion mobility spectrometry.

Ion mobility spectrometry, originally used to detect chemical warfare agents, explosives and illegal drugs, is now frequently applied in the field of process analytics. The method combines both high sensitivity (detection limits down to the ng to pg per liter and ppb(v)/ppt(v) ranges) and relatively low technical expenditure with a high-speed data acquisition. In this paper, the working principles of IMS are summarized with respect to the advantages and disadvantages of the technique. Different ionization techniques, sample introduction methods and preseparation methods are considered. Proven applications of different types of ion mobility spectrometer (IMS) used at ISAS will be discussed in detail: monitoring of gas insulated substations, contamination in water, odoration of natural gas, human breath composition and metabolites of bacteria. The example applications discussed relate to purity (gas insulated substations), ecology (contamination of water resources), plants and person safety (odoration of natural gas), food quality control (molds and bacteria) and human health (breath analysis).