Exhaled Breath Condensate Analysis Research Articles

A wearable exhaled breath condensate (EBC) collector with controllable condensation microfluidics and a branched hydrophilic film

In previous research, exhaled breath condensate (EBC) analysis has emerged as a promising noninvasive method for assessing human health, potentially increasing patient testing acceptance. However, current EBC collection devices require cooling equipment for temperature reduction, leading to issues such as increased size, increased energy consumption, and a lack of real-time collection capability. In this study, an EBC collector was developed that integrates controllable condensation microfluidics with a biomimetic water-collecting film. Microfluids induce an endothermic reaction by rapidly dissolving water and NH4NO3 within the chamber, reducing the surface temperature to 3.5 °C. When positioned inside a mask, exhaled air undergoes condensation and collection on the branched biomimetic film. Human tests were conducted to analyze caffeine and nicotine in the collected EBC samples using LC–MS, demonstrating the device’s advantages in terms of power-free operation, wearable design, and rapid sample collection for metabolite detection.

The optimization of low volume-SPME method for volatilomics analysis of exhaled breath condensate

The analysis of breath can provide insight into the metabolic state of the body. However, the collection, transport, and storage of breath samples present certain challenges. The collection of exhaled breath condensate (EBC) emerges as a promising method for breath analysis. Despite this, the volume of EBC is relatively low, necessitating a sensitive method capable of extracting analytes from a small sample volumes. Therefore, this study aimed to develop and optimize a robust and green solid-phase extraction method for the analysing volatile organic compounds (VOCs) of various polarities in a low-volume EBC using a statistical design of experiment (DoE).Based on the screening experiments and DoE, the optimal conditions were determined to be 15 min equilibration time, and 20 min of extraction at 44 °C using DVB/CAR/PDMS fibre in direct immersion (DI) mode. The method was validated for linearity, precision and accuracy, yielding satisfactory results. The optimized method was applied to quantify the concentration of selected VOCs in real samples collected from a healthy subject, as well as after consumption of coffee and smoking of an electronic cigarette. Many of the selected compounds are associated with diseases and are challenging to detect in specimens from a healthy subject, but this method can measure them as potential disease markers.The proposed method enables the analysis of VOCs is very low-volume samples (200 µL), which is particularly important for studies involving body fluids. Notably, the applied approach demonstrated that different VOCs, representing various chemical classes and polarities, can be detected and quantified.

Measurement of Human and Bovine Exhaled Breath Condensate pH Using Polyaniline-Modified Flexible Inkjet-Printed Nanocarbon Electrodes.

The collection, processing, and electrochemical analysis of exhaled breath condensate (EBC) from healthy human and animal subjects is reported on. EBC is a biospecimen potentially rich in biomarkers of respiratory disease. The EBC pH was analyzed potentiometrically using a disposable polyaniline (PANI)-modified inkjet-printed (IJP) carbon electrode. Comparison measurements were performed using a commercial screen-printed carbon (SPC) electrode. The PANI-modified electrodes exhibited reproducible and near-Nernstian responses for pH values between 2 and 9 with slopes from -50 to -60 mV/dec. The PANI-modified IJP carbon electrode exhibited a faster response time and superior reproducibility to the modified SPC electrode. In proof-of-concept studies, the healthy human EBC pH was found to be 6.57 ± 0.09 and the healthy bovine EBC pH was 5.9 ± 0.2. All pH determined using the PANI-modified electrodes were in good agreement with the pH determined using a micro glass pH electrode. An RTube device was used to collect EBC from humans while a modified device was used to collect EBC from calves in the field. EBC volumes of 0.5-2 mL for 5-6 min of tidal breathing were collected from healthy animals. The pH of EBC from healthy calves (17 animals) depends on their age from 1 to 9 weeks with values ranging from 5.3 to 7.2. A distinct alkaline shift was observed for many animals around 20 days of age. The bovine EBC pH also depends on the ambient temperature and humidity at the time of collection. The results indicate that the PANI-modified IJP carbon electrodes outperform commercial SPC and provide reproducible and accurate measurement of pH across various biospecimen types.

A smart mask for exhaled breath condensate harvesting and analysis.

Recent respiratory outbreaks have garnered substantial attention, yet most respiratory monitoring remains confined to physical signals. Exhaled breath condensate (EBC) harbors rich molecular information that could unveil diverse insights into an individual's health. Unfortunately, challenges related to sample collection and the lack of on-site analytical tools impede the widespread adoption of EBC analysis. Here, we introduce EBCare, a mask-based device for real-time in situ monitoring of EBC biomarkers. Using a tandem cooling strategy, automated microfluidics, highly selective electrochemical biosensors, and a wireless reading circuit, EBCare enables continuous multimodal monitoring of EBC analytes across real-life indoor and outdoor activities. We validated EBCare's usability in assessing metabolic conditions and respiratory airway inflammation in healthy participants, patients with chronic obstructive pulmonary disease or asthma, and patients after COVID-19 infection.

Assessing the inhaled dose of nanomaterials by nanoparticle tracking analysis (NTA) of exhaled breath condensate (EBC) and its relationship with lung inflammatory biomarkers

The widespread and increasing use of nanomaterials has resulted in a higher likelihood of exposure by inhalation for nanotechnology workers. However, tracking the internal dose of nanoparticles deposited at the airways level, is still challenging.To assess the suitability of particle number concentration determination as biomarker of internal dose, we carried out a cross sectional investigation involving 80 workers handling nanomaterials. External exposure was characterized by portable counters of particles DISCminiTM (Testo, DE), allowing to categorize 51 workers as exposed and 29 as non-exposed (NE) to nanoparticles. Each subject filled in a questionnaire reporting working practices and health status. Exhaled breath condensate was collected and analysed for the number of particles/ml as well as for inflammatory biomarkers.A clear-cut relationship between the number of airborne particles in the nano-size range determined by the particle counters and the particle concentration in exhaled breath condensate (EBC) was apparent. Moreover, inflammatory cytokines (IL-1β, IL-10, and TNF-α) measured in EBC, were significantly higher in the exposed subjects as compared to not exposed. Finally, significant correlations were found between external exposure, the number concentration of particles measured by the nanoparticle tracking analysis (NTA) and inflammatory cytokines. As a whole, the present study, suggests that NTA can be regarded as a reliable tool to assess the inhaled dose of particles and that this dose can effectively elicit inflammatory effects.

Analysis of exhaled breath condensate in patients with asthma and recurrent wheezing

Wheezing is the most common clinical symptom of bronchial obstructive syndrome. The functions of pulmonary surfactant (PS) depend directly on the level of total phospholipids (TP). This can be used to assess the integrity of the cell membrane structure. Objective. To determine the level of total phospholipids and calcium in exhaled breath condensate in young children with recurrent wheezing and asthma. Materials and methods. The study included 77 patients divided into 3 groups. Group 1 included 30 patients (20 girls and 10 boys) with no more than 2 episodes of recurrent wheezing in their lifetime; Group 2 -10 patients (6 boys and 4 girls) with more than 3 episodes of recurrent wheezing, and 37 patients (15 girls, 22 boys) with asthma. The control group consisted of 20 conditionally healthy children (11 girls, 9 boys). All patients were aged from 3 months to 6 years. The determination of phospholipid levels and calcium (Ca) in the exhaled breath condensate was performed in dynamics - during the period of clinical manifestations and remission. Results. The level of phospholipids in the exhaled breath condensate was the highest in patients of all groups at the stage of clinical manifestations compared with the period of remission, while its lowest level was noted in the control group. During the peak of the disease, the level of Ca in the exhaled breath condensate was significantly lower compared to the period of remission. The control group had the highest level. Conclusions. The exhaled breath condensate's high level of phospholipids in the first days of the disease confirms cell damage in the presence of inflammation. Additionally, the exhaled breath condensate's Ca level is lowest during this period, possibly indicating its involvement in cell damage

Advances in proteomics methods for the analysis of exhaled breath condensate.

The analysis of exhaled breath condensate (EBC) demonstrates a promising avenue of minimally invasive biopsies for diagnostics. EBC is obtained by cooling exhaled air and collecting the condensation to be utilized for downstream analysis using various analytical methods. The aqueous phase of breath contains a large variety of miscible small compounds including polar electrolytes, amino acids, cytokines, chemokines, peptides, small proteins, metabolites, nucleic acids, and lipids/eicosanoids-however, these analytes are typically present at minuscule levels in EBC, posing a considerable technical challenge. Along with recent improvements in devices for breath collection, the sensitivity and resolution of liquid chromatography coupled to online mass spectrometry-based proteomics has attained subfemtomole sensitivity, vastly enhancing the quality of EBC sample analysis. As a result, proteomics analysis of EBC has been expanding the field of breath biomarker research. We present an au courant overview of the achievements in proteomics of EBC, the advancement of EBC collection devices, and the current and future applications for EBC biomarker analysis.

Biosensor for Rapid Measurement of Lactate in Exhaled Breath Condensate

Lactate concentration plays an important role in disease diagnostics. Higher lactate concentrations are associated with sepsis and septic shock [2], trauma [3], tissue hypoxia due to acute lung injury [4], and respiratory diseases [5]. Lactate level is also widely used in monitoring athletes’ training and fitness as long-term exhausting exercise leads to a rapid increase in lactate level and result in muscle weakening and fatigue [6]. Compared with the traditional blood sample media which is invasive and capillary blood monitoring is not suitable for routine clinical use, the exhaled breath condensate (EBC) is simple and non-invasive method with no potential for adverse effects for lactate analysis.In this presentation, a point-of-care electrochemical biosensor is introduced for the EBC lactate analysis. This disposable and low-cost sensor is aimed to be inserted in an EBC collection device developed by our team that enables real-time monitoring and rapid measurements of EBC lactate level. The sensor measurement is done directly after EBC sample collection, that minimizes the risk of sample degradation and is cheaper than the traditional laboratory techniques such as fluorometry or mass spectrometry.The sensor is modified with poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS), Prussian blue (PB) and lactate oxidase (LOD) enzyme. Lactate concentration is measured from amperometric current response generated by the redox reactions on sensor surface. The enzyme gel formulation is optimized based on rotating disk electrode (RDE) experiments and kinetics analysis, investigating the effect of LOD concentration and gel thickness, and was then applied to disposable sensors.Finally, human EBC analysis is conducted on healthy subjects at rest and after 30 min of intense aerobic cycling exercise. Sensor’s results in real EBC lactate measurements demonstrate a good correlation with fluorometry and mass spectrometry calibrations and exhibit high stability and sensitivity in both short-term and long-term use. Mass production of the sensor is done by robotic gel dispensing using a Biodot aspirate dispensing machine.Note: This presentation is based on the corresponding author's recent publication ‘Rapid Measurement of Lactate in the Exhaled Breath Condensate: Biosensor Optimization and In-Human Proof of Concept’ in ACS Sensors (doi.10.1021/acssensors.2c01739) [1]

MicroRNAs in exhaled breath condensate for diagnosis of lung cancer in a resource-limited setting: a concise review.

Lung cancer is one of the common cancers globally with high mortality and poor prognosis. Most cases of lung cancer are diagnosed at an advanced stage due to limited diagnostic resources. Screening modalities, such as sputum cytology and annual chest radiographs, have not proved sensitive enough to impact mortality. In recent years, annual low-dose computed tomography has emerged as a potential screening tool for early lung cancer detection, but it may not be a feasible option for developing countries. In this context, exhaled breath condensate (EBC) analysis has been evaluated recently as a noninvasive tool for lung cancer diagnosis. The breath biomarkers also have the advantage of differentiating various types and stages of lung cancer. Recent studies have focused more on microRNAs (miRNAs) as they play a key role in tumourigenesis by regulating the cell cycle, metastasis and angiogenesis. In this review, we have consolidated the current published literature suggesting the utility of miRNAs in EBC for the detection of lung cancer.

Mechanisms Contributing to the Comorbidity of COPD and Lung Cancer.

Lung cancer and chronic obstructive pulmonary disease (COPD) often co-occur, and individuals with COPD are at a higher risk of developing lung cancer. While the underlying mechanism for this risk is not well understood, its major contributing factors have been proposed to include genomic, immune, and microenvironment dysregulation. Here, we review the evidence and significant studies that explore the mechanisms underlying the heightened lung cancer risk in people with COPD. Genetic and epigenetic changes, as well as the aberrant expression of non-coding RNAs, predispose the lung epithelium to carcinogenesis by altering the expression of cancer- and immune-related genes. Oxidative stress generated by tobacco smoking plays a role in reducing genomic integrity, promoting epithelial-mesenchymal-transition, and generating a chronic inflammatory environment. This leads to abnormal immune responses that promote cancer development, though not all smokers develop lung cancer. Sex differences in the metabolism of tobacco smoke predispose females to developing COPD and accumulating damage from oxidative stress that poses a risk for the development of lung cancer. Dysregulation of the lung microenvironment and microbiome contributes to chronic inflammation, which is observed in COPD and known to facilitate cancer initiation in various tumor types. Further, there is a need to better characterize and identify the proportion of individuals with COPD who are at a high risk for developing lung cancer. We evaluate possible novel and individualized screening strategies, including biomarkers identified in genetic studies and exhaled breath condensate analysis. We also discuss the use of corticosteroids and statins as chemopreventive agents to prevent lung cancer. It is crucial that we optimize the current methods for the early detection and management of lung cancer and COPD in order to improve the health outcomes for a large affected population.

Diet Quality and Exhaled Breath Condensate Markers in a Sample of School-Aged Children.

Exhaled breath condensate (EBC) analysis is a recently developed, non-invasive method used to identify and quantify biomarkers, mainly those coming from the lower respiratory tract. It seems that diet can influence the airway's inflammation and change the exhaled breath composition. This study aimed to assess the association between diet quality intake and markers in EBC among school-aged children. A cross-sectional analysis included 150 children (48.3% females, aged 7-12 years, mean age: 8.7 ± 0.8 years) from 20 schools across Porto, Portugal. We assessed diet quality through the Healthy Eating Index (HEI)-2015, which was estimated based on a single 24 h food recall questionnaire. EBC samples were collected, and we assessed their ionic content (Na+, K+) and conductivity. The association between diet quality and Na+, K+, Na+/K+ ratio and conductivity was estimated using logistic regression models adjusted for potential confounders. After adjustment, a higher quality diet score increases the odds of higher conductivity values of the EBC (aOR = 1.04, 95%CI 1.00; 1.08). Our findings suggest that a higher diet quality in school-aged children is associated with higher conductivity levels of the EBC.

A Mycobacterium tuberculosis fingerprint in human breath allows tuberculosis detection

An estimated one-third of tuberculosis (TB) cases go undiagnosed or unreported. Sputum samples, widely used for TB diagnosis, are inefficient at detecting infection in children and paucibacillary patients. Indeed, developing point-of-care biomarker-based diagnostics that are not sputum-based is a major priority for the WHO. Here, in a proof-of-concept study, we tested whether pulmonary TB can be detected by analyzing patient exhaled breath condensate (EBC) samples. We find that the presence of Mycobacterium tuberculosis (Mtb)-specific lipids, lipoarabinomannan lipoglycan, and proteins in EBCs can efficiently differentiate baseline TB patients from controls. We used EBCs to track the longitudinal effects of antibiotic treatment in pediatric TB patients. In addition, Mtb lipoarabinomannan and lipids were structurally distinct in EBCs compared to ex vivo cultured bacteria, revealing specific metabolic and biochemical states of Mtb in the human lung. This provides essential information for the rational development or improvement of diagnostic antibodies, vaccines and therapeutic drugs. Our data collectively indicate that EBC analysis can potentially facilitate clinical diagnosis of TB across patient populations and monitor treatment efficacy. This affordable, rapid and non-invasive approach seems superior to sputum assays and has the potential to be implemented at point-of-care.

Rapid Measurement of Lactate in the Exhaled Breath Condensate: Biosensor Optimization and In-Human Proof of Concept.

Lactate concentration is of increasing interest as a diagnostic for sepsis, septic shock, and trauma. Compared with the traditional blood sample media, the exhaled breath condensate (EBC) has the advantages of non-invasiveness and higher user acceptance. An amperometric biosensor was developed and its application in EBC lactate detection was investigated in this paper. The sensor was modified with PEDOT:PSS-PB, and two different lactate oxidases (LODs). A rotating disk electrode and Koutecky-Levich analysis were applied for the kinetics analysis and gel optimization. The optimized gel formulation was then tested on disposable screen-printed sensors. The disposable sensors exhibited good performance and presented a high stability for both LOD modifications. Finally, human EBC analysis was conducted from a healthy subject at rest and after 30 min of intense aerobic cycling exercise. The sensor coulometric measurements showed good agreement with fluorometric and triple quadrupole liquid chromatography mass spectrometry reference methods. The EBC lactate concentration increased from 22.5 μM (at rest) to 28.0 μM (after 30 min of cycling) and dropped back to 5.3 μM after 60 min of rest.

High-resolution metabolomics-based biomarker discovery using exhaled breath condensate from patients with lung cancer

Early diagnosis and treatment are critical for improving the survival of patients with lung cancer, which is the leading cause of cancer-related deaths worldwide. In this study, we investigated whether the metabolomics analysis of exhaled breath condensate (EBC) from patients with lung cancer can provide biomarkers that can be used for noninvasive screening for lung cancer diagnosis. EBC samples obtained from patients with lung cancer (n = 20) and healthy individuals (n = 5) were subjected to high-resolution metabolomics (HRM) using liquid chromatography–mass spectrometry (LC–MS). Univariate analysis, with a false discovery rate (FDR), q = 0.05, and hierarchical clustering analysis were performed to discover significantly different metabolites between the healthy controls and patients with lung cancer. This was followed by the identification of the metabolites using the METLIN database. Pathway analysis based on the identified metabolites revealed that arachidonic acid (AA) metabolism was the most significantly affected pathway. Finally, 5-hydroxyicosatetraenoic acid (HETE) (m/z 343.2233, [M + Na]+), a metabolite involved in AA metabolism, was found to be significantly higher in patients with lung cancer than in healthy counterparts. Our finding suggested that the HRM of EBC samples is a useful approach for identifying biomarkers for noninvasive screening for lung cancer diagnosis.

Lyophilization with Subsequent Derivatization vs Microextraction by Packed Sorbent (MEPS) in the Analysis of Exhaled Breath Condensate by Gas Chromatography–Mass Spectrometry

Lyophilization with Subsequent Derivatization vs Microextraction by Packed Sorbent (MEPS) in the Analysis of Exhaled Breath Condensate by Gas Chromatography–Mass Spectrometry

Exhaled Breath Condensate Analysis in Chronic Obstructive Pulmonary Disease

The increasing focus on airway inflammation in the pathogenesis of chronic obstructive pulmonary disease (COPD) has led to development and evolution of tools to measure it. Direct assessment of airway inflammation requires invasive procedures, and hence, has obvious limitations. Non-invasive methods to sample airway secretions and fluids offer exciting prospects. Analysis of exhaled breath condensate (EBC) is rapidly emerging as a novel non-invasive approach for sampling airway epithelial lining fluid and offers a convenient tool to provide biomarkers of inflammation. It has definite advantages that make it an attractive and a feasible option. It is a source of mediators and molecules that are the causes or consequences of the inflammatory process. Measurement of such markers is increasingly being explored for studying airway inflammation qualitatively and quantitatively in research studies and for potential clinical applications. These biomarkers also have the potential to develop into powerful research tools in COPD for identifying various pathways of pathogenesis of COPD that may ultimately provide specific targets for therapeutic intervention. The EBC analysis is still an evolving noninvasive method for monitoring of inflammation and oxidative stress in the airways. The limited number of studies available on EBC analysis in COPD have provided useful information although definite clinical uses are yet to be defined. Evolving technologies of genomics, proteomics, and metabonomics may provide deeper and newer insights into the molecular mechanisms underlying the pathogenesis of COPD.

Abstract 2797: Proteomic signature in exhaled breath condensates for a non-invasive diagnostics of lung cancer

Abstract Because of the invasiveness of examination methods as bronchoscopy for the diagnosis of lung cancer, there is currently no preventive screening for this disease. Thus, lung cancer is usually diagnosed in stage 3 or 4, when it is too late for successful treatment. However, if diagnosed early, it can be cured entirely. Therefore, it is necessary to find a suitable method to screen the high-risk populations and reveal lung cancer in the early stages. Since the mucous membranes of the respiratory tract are naturally moistened by fluids containing proteins, metabolites, chemoattractants and growth factors released by the highly active epithelium, the collection of the respiratory tract fluids can be used to diagnose malignant and benign respiratory tract diseases. The most commonly used method in this sense is bronchoalveolar lavage. However, the technique itself is not pleasant and can be accompanied by possible complications. Therefore, intensive research work is conducted to develop non-invasive methods for biomarker detection in exhaled breath, sputum, saliva or nasal secretion. We have studied an exhaled breath condensate (EBC) matrix, which is collected non-invasively and the patients are not suffering from its collection. Several studies have reported proteomic analysis of EBC while identifying low hundreds of proteins by mass spectrometry (MS). We have developed a potent gel-free MS-based approach for sample preparation and analysis. Together with a powerful search tool, we were able to identify a ten times higher number of proteins across a comparable group of individuals of about three hundred peoples’ samples. Our approach seems to be highly reproducible and applicable to various respiratory and systemic diseases. In our work focused on non-small lung cancer biomarkers compared to chronic obstructive pulmonary disease (COPD) and healthy controls, we have identified 7694 proteins and of them, 6525 proteins were quantified at least in one replicate across 296 individuals’ samples measured in triplicates. Combining univariate and multivariate statistical approaches and sensitivity analysis, we have suggested biomarkers that could distinguish lung cancer patients from COPD and healthy individuals. This research was supported by grants from the Czech Ministry of Education, Youth and Sports (EATRIS-CZ - LM2018133), European Regional Development Fund - Project ENOCH (No. CZ.02.1.01/0.0/0.0/16_019/0000868) and IGA_LF_2021_036 (Palacky University in Olomouc). Citation Format: Jana Vaclavkova, Jana Vrbkova, Pavla Kourilova, Dusan Holub, Juraj Kultan, Petr Jakubec, Ondrej Fischer, Frantisek Kopriva, Vendula Latalova, Tatiana Gvozdiakova, Marian Hajduch, Petr Dzubak. Proteomic signature in exhaled breath condensates for a non-invasive diagnostics of lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2797.

Comparative proteomic analysis of exhaled breath condensate between lung adenocarcinoma and CT-detected benign pulmonary nodule patients.

Lung cancer is the leading cause of cancer mortality worldwide. The collection of exhaled breath condensate (EBC) is a non-invasive method that may have enormous potential as a biomarker for the early detection of lung cancer. To investigate the proteomic differences of EBC between lung cancer and CT-detected benign nodule patients, and determine whether these proteins could be potential biomarkers. Proteomic analysis was performed on individual samples from 10 lung cancer patients and 10 CT-detected benign nodule patients using data-independent acquisition (DIA) mass spectrometry. A total of 1,254 proteins were identified, and 21 proteins were differentially expressed in the lung adenocarcinoma group compared to the benign nodule group (p< 0.05). The GO analysis showed that most of these proteins were involved in neutrophil-related biological processes, and the KEGG analysis showed these proteins were mostly annotated to pyruvate and propanoate metabolism. Through protein-protein interactions (PPIs) analysis, ME1 and LDHB contributed most to the interaction-network of these proteins. Significantly differentially expressed proteins were detected between lung cancer and the CT-detected benign nodule group from EBC samples, and these proteins might serve as potential novel biomarkers of EBC for early lung cancer detection.

Challenges in Quantifying 8-OHdG and 8-Isoprostane in Exhaled Breath Condensate.

Exhaled breath condensate (EBC) has attracted substantial interest in the last few years, enabling the assessment of airway inflammation with a non-invasive method. Concentrations of 8-Hydroxydesoxyguanosine (8-OHdG) and 8-isoprostane in EBC have been suggested as candidate biomarkers for lung diseases associated with inflammation and oxidative stress. EBC is a diluted biological matrix and consequently, requires highly sensitive chemical analytic methods (picomolar range) for biomarker quantification. We developed a new liquid chromatography coupled to tandem mass spectrometry method to quantify 8-OHdG and 8-isoprostane in EBC simultaneously. We applied this novel biomarker method in EBC obtained from 10 healthy subjects, 7 asthmatic subjects, and 9 subjects with chronic obstructive pulmonary disease. Both biomarkers were below the limit of detection (LOD) despite the good sensitivity of the chemical analytical method (LOD = 0.5 pg/mL for 8-OHdG; 1 pg/mL for 8-isoprostane). This lack of detection might result from factors affecting EBC collections. These findings are in line with methodological concerns already raised regarding the reliability of EBC collection for quantification of 8-OHdG and 8-isoprostane. Precaution is therefore needed when comparing literature results without considering methodological issues relative to EBC collection and analysis. Loss of analyte during EBC collection procedures still needs to be resolved before using these oxidative stress biomarkers in EBC.

Metabolomics Diagnosis of COVID-19 from Exhaled Breath Condensate.

Infection from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can lead to severe respiratory tract damage and acute lung injury. Therefore, it is crucial to study breath-associated biofluids not only to investigate the breath’s biochemical changes caused by SARS-CoV-2 infection, but also to discover potential biomarkers for the development of new diagnostic tools. In the present study, we performed an untargeted metabolomics approach using a bidimensional gas chromatography mass spectrometer (GCxGC-TOFMS) on exhaled breath condensate (EBC) from COVID-19 patients and negative healthy subjects to identify new potential biomarkers for the noninvasive diagnosis and monitoring of the COVID-19 disease. The EBC analysis was further performed in patients with acute or acute-on-chronic cardiopulmonary edema (CPE) to assess the reliability of the identified biomarkers. Our findings demonstrated that an abundance of EBC fatty acids can be used to discriminate COVID-19 patients and that they may have a protective effect, thus suggesting their potential use as a preventive strategy against the infection.