Table 1.
Qualitative synthesis of articles that underwent data extraction.
| Author, Year, Country | Study Objectives | Population Studied and Number of Participants | Control Population | Method of EBC Collection and Analysis | Main Findings | Quality Score |
|---|---|---|---|---|---|---|
| Aksu, 2012, Turkey [23] | To investigate lower airway inflammation using malondialdehyde and total protein measurement in exhaled breath condensate | 12 Mild asthma, 53 persistent rhinitis, and 16 concomitant asthma and rhinitis | 13 Control subjects (5 males and 8 females) | EBC was collected with a homemade apparatus for max 15 min at 10.00 am, no nose clip, immediately stored at −80 °C | MDA and total protein levels in EBC did not differ among the groups | 14 Moderate |
| Andreoli, 2003, Italy [24] | Method development | None | 2 Non-smoking and 13 asymptomatic smoking control subjects | EBC collected on EcoScreen, 15 min with nose clip, and then quickly stored at −80 °C | MDA concentrations were consistent with those determined in EBC using an independent HPLC-FLD method, mass spectrometry to EBC analysis could play a fundamental role in the validation of EBC as a new accessible biological matrix and in the definition of standardized EBC collection | 12 Moderate |
| Antus, 2015, Hungary [25] | To investigate oxidative stress in cystic fibrosis (CF) patients | 40 Patients with CF | 25 Control subjects (13 males and 12 females) |
EBC collection with an EcoScreen condenser using MDA reagent kit, MDA concentrations in EBC were measured with an isocratic HPLC system | 19 High | |
| Antus, 2013, Hungary [26] | To investigate MDA levels in patients with acute COPD exacerbation | 55 Patients with COPD | 20 Healthy controls, ex-smokers (10 females and 10 males) | EBC collected for 10 min EcoScreen condenser, samples were stored frozen at −80 °C before analysis, isocratic HPLC system using MDA reagent kit | 12 Moderate | |
| Antczak, 1997, Poland [27] | To determine whether asthmatic patients exhale more H2O2 and TBARS than healthy subjects | 21 Asthmatic patients | 10 Healthy subjects (6 males and 4 females) | EBC collected in a tube installed in a polystyrene foam container filled with ice and salt, for 20 min, and aliquotes were transferred in Eppendorf tubes stored at −80 °C for no longer than 7 days until measurement, TBAR species in expired breath condensate was determined according to the method of YAGI | H2O2 was 26 times higher in asthmatic EBC as compared with TBARS, these findings were statistically significant | 15 Moderate |
| Araneda, 2005, Chile [28] | To investigate lung oxidative stress in subjects exercising in moderate and high altitude | None | 6 Healthy males | EBC collected with self-constructed device for 20 min, condensated at −5 °C, and stored in liquid nitrogen, MDA was determined with HPLC according to Larstad | Lung OS may constitute a pathogenic factor in acute mountain sickness | 11 Moderate |
| Araneda, 2012, Chile [29] | To investigate the impact of an endurance race on pulmonary pro-oxidative formation and lipoperoxidation | 41 Healthy recreational runners divided into 3 groups according to the races they decided to run (10, 21.1, and 41.2 km) non-smokers | Two groups (10 and 42.2 km) with MDA in EBC, pre-race data was available just for these subjects | EBC collected with self-constructed device for 10 to 20 min, with nose clip, condensated at −5 °C, and stored in liquid nitrogen, MDA was determined with HPLC according to Larstad | Intense prolonged exercise favors an increase in pulmonary pro-oxidative levels, with no modifications on lipoperoxidation; Running time relates to the magnitude of acute post exercise pro-oxidative formation | 12 Moderate |
| Araneda, 2013, Chile [30] | To investigate pro-oxidative, lipid peroxidation, and inflammation in EBC, in subjects running 10 km | None | 10 Physically active healthy subjects (9 males and 1 female), EBC measured 20 min before the race | EBC collected with self-constructed device for 10 to 15 min, with nose clip, condensated at −5 °C, and stored in liquid nitrogen, MDA was determined with HPLC according to Larstad | Unlike the previous results obtained in amateur runners, in physically active subjects, 10 km produces an increase in oxygen- and nitrogen-derived pro-oxidative species, without early peroxidation | 7 Low |
| Barregard, 2007, Sweden [31] | To examine whether short-term exposure to wood smoke in healthy subjects affects markers of pulmonary inflammation and oxidative stress | 13 Healty subjects (6 males and 7 females), EBC measured prior to exposure | EBC was collected with an Rtube, with nose clip, until 80 L were collected, samples were frozen at −20 °C, MDA was determined with HPLC according to Larstad | Wood smoke exposure affects the respiratory tract, especially the lower airways | 10 Moderate | |
| Bartoli, 2011, Italy [32] | To assess the usefulness of MDA in EBC in different groups of pulmonary diseases | 64 Subjects with asthma, 19 with bronchiectasies, 73 with COPD, 38 with idiopathic pulmonary fibrosis | 14 Healthy non-smoking subjects | EBC was collected on an EcoScreen device, during 15 min, samples were immediately stored at −80 °C and analyzed within 6 months, with HPLC according to Larstad | Subjects with chronic airway disorders have increased levels of MDA in EBC, MDA concentrations in EBC are related to FEV1 and neutrophilic inflammation, particularly in COPD patients. | 15 Moderate |
| Brand, 2013, Germany [33] | To investigate if short-term exposure to welding fumes results in changes in lung function and early stages of inflammatory reactions | 12 Healthy non-smoking males | EBC was collecte on an ECOSCREEN device wearing a nose clip, for 20 min, samples stored at −80 °C until analysis, MDA was derivatized with DNPH, and then separated by HPLC and determined by mass spectrometry | In healthy, young subjects, neither changes in spirometry nor changes in inflammatory markers measured in exhaled breath condensate could be detected after short-term exposure | 12 Moderate | |
| Caglieri, 2006, Italy [34] | To investigate chromium levels in EBC of workers exposed to Cr(VI) and to assess their relationship with biochemical changes in the airways by analyzing EBC biomarkers of oxidative stress | 24 Chrome-plating workers | 25 Control subjects (13 males and 12 females, 5 ex-smokers and 20 non-smokers) | EBC was collected on TURBO DECCS, condensation temperature of −5 °C, during 15 min; MDA-EBC was measured by tandem liquid chromatography–mass spectrometry | Cr-EBC levels correlated with those of H2O2-EBC and MDA-EBC, as well as with urinary Cr levels. | 14 Moderate |
| Casimirri, 2015, Italy [35] | Casimirri, 2015, Italy | Casimirri, 2015, Italy | Casimirri, 2015, Italy | Casimirri, 2015, Italy | Professional exposure to chlorinated agents increases EBC biomarkers of oxidativestress and inflammation | 15 Moderate |
| Corradi, 2003, Italy [36] | To evaluate if aldehydes could be measured in EBC, to assess the influence of sampling procedures, to compare levels of different pulmonary disease groups with those of a control group | 20 Patients with stable COPD (18 males and 2 females) | 12 Smoking (9 males and 3 females) and 20 non-smoking (17 males and 3 females) control subjects | EBC was collected on a Tygon Tube immersed in thawing ice, and then frozen to −80 °C, no nose clip, MDA measured with LC-MS tandem | Aldehydes were identified in EBC, all, but were lower in control groups | 16 Moderate |
| Corradi, 2002, Italy [37] | To investigate if short-term exposure to ozone (O3) induces changes in biomarkers of lung inflammation and oxidative stress in EBC | 22 Non-smoking healthy control subjects (12 males and females) | EBC was collected on an ECO SCREEN device, during 15 min, frozen at −80 °C, MDA was measured as TBARS according to Nowak | A single 2-hour exposure to 0.1 ppm of O3 induces changes in biomarkers of inflammation and oxidative stress in those susceptible | 11 Moderate | |
| Corradi, 2004, Italy [38] | To compare aldehyde levels resulting from lipid peroxidation in EBC and induced sputum (IS) supernatant of subjects with asthma and chronic obstructive pulmonary disease | 21 Subjects with COPD, 10 asthmatics | 9 Healthy non-smoking control subjects (8 females and 1 male) | EBC was collected on a two glass chamber device from Incofar, no nose clip for 20 min, samples were stored at −80 °C, and MDA measured according to Larstad | Aldehydes can be detected in both exhaled breath condensate and supernatant of induced sputum, but their relative concentrations are different and not correlated with each other | 12 Moderate |
| Doruk, 2011, Turkey [39] | To investigate oxidative stress in the lungs associated with tobacco smoke and to evaluate the effect of this stress with pulmonary function tests | 69 Healthy subjects divided into 3 groups according to their exposure to tobacco smoke: 26 current smokers (23 males and 3 females), 21 subjects (15 males and 6 females) who did not smoke within the last year but had second-hand smoking and 22 (100 males and 12 females) with no tobacco smoke exposure | EBC was collected on an ECOSCREEN device for 15 min, wearing a nose clip, samples stored at −70 °C, MDA measured as TBARS | The levels of MDA, 8-OHdG, SOD, and GSH-Px were higher in smokers; NO levels gradually increased from Group I to Group III; MDA levels were lower in Group III than Group II | 14 Moderate | |
| Goldoni, 2004, Italy [40] | To investigate whether EBC can be used as a suitable matrix to assess target tissue dose and effects of inhaled cobalt and tungsten, using EBC malondialdehyde (MDA) as a biomarker of pulmonary oxidative stress | 33 Workers exposed to Co and W in workshops producing either diamond tools or hard-metal mechanical parts | 16 Control subjects (11 males and 5 females) | EBC was collected on a homemade apparatus, during 10 min, samples were transported in ice to the lab, and then stored at −80 °C, MDA was measured by liquid chromatography–tandem mass spectrometry (LC-MS/MS) | MDA levels were increased depending on cobalt concentration and were enhanced by coexposure to tungsten. | 11 Moderate |
| Goldoni, 2013, Italy [41] | To compare the concentration of several biomarkers in whole (W-EBC) and fractionated EBC (A-EBC) | 45 Healthy control subjects (10 males and 35 females, 6 smokers and 39 non-smokers) | EBC was collected on a homemade apparatus for the fractioned EBC and a TURBODECCS for the whole EBC for 15 min without a nose clip, EBC was centrifuged, and then stored at −80 °C, MDA was measured by liquid chromatography–tandem mass spectrometry | H2O2, 8-isoprostane, malondialdehyde, and 4-hydroxy-2-nonhenal were all higher in W-EBC, suggesting a contribution from the upper airways to oxidative stress biomarkers in apparently healthy subjects | 10 Moderate | |
| Goldoni, 2005, Italy [42] | To test the effect of condensation temperature on the parameters of exhaled breath condensate and the levels of selected biomarkers | 24 Healthy control subjects (13 males and 11 females, 3 ex-smokers and 21 non-smokers) | EBC was collected on a TURBODECCS, during 10 min, at different temperatures, samples were centrifuged, and then stored at −80 °C, MDA was measured by liquid chromatography–tandem mass spectrometry (LC-MS/MS) within 2 weeks | Cooling temperature of exhaled breath condensate collection influenced selected biomarkers | 13 Moderate | |
| Gong, 2014, USA/China [43] | To compare ultrafine particles (UFPs) and fine particles (PM2.5) with respect to their associations with biomarkers reflecting multiple pathophysiological pathways linking exposure and cardiorespiratory events | 125 Healthy non-smoking individuals (initially 64 males and 64 females) working on a campus near Peking’s University Health Sciences Centre | EBC was collected on an ECOSCREEN device, collected for 20 min, with nose clip, aliquotes stored at −70 °C, MDA was measured as MDA-TBA in HPLC | Associations of certain biomarkers with UFPs had different lag patterns as compared with those with PM2.5, suggesting that the ultrafine size fraction and the fine size fraction of PM2.5 are likely to affect PM-induced pathophysiological pathways independently | 15 Moderate | |
| Graczyk, 2016, Switzerland [44] | To investigate time course changes of particle-associated oxidative stress in exposed tungsten inert gas welders | 20 Non-smoking healthy volunteers, with less than 1 year of apprentice in welding | EBC was collected on an Rtube, during 10 min, while wearing a nose clip, EBC samples were stored at −70 °C, MDA was measured by HPLC separation and fluorescence detection TBARS | A 60-minute exposure to TIG welding fume in a controlled, well-ventilated setting induced acute oxidative stress at 3 h post exposure in healthy, non-smoking apprentice welders not chronically exposed to welding fumes | 10 Moderate | |
| Gube, 2010, Germany [45] | To investigate the effect of welding as well as the impact of smoking and protection measures on biological markers in EBC | 45 Male welders | 24 Healthy males, non-exposed | EBC was collected on an ECOSCREEN device, wearing a nose clip, as long as 200 L of exhaled breath was collected, then stored at −80 °C, MDA was measued with a previously described but slightly changed method, i.e., MDA derivatized with DNPH and separated by HPLC, and then tandem mass spectrometry | Welders showed significantly increased concentrations of all these parameters at baseline as compared with non-exposed controls | 14 Moderate |
| Inonu, 2012, Turkey [46] | To evaluate the differences in the burden of oxidative stress in patients with COPD, smokers, and non-smokers by measuring hydrogen peroxide (H2O2), malondialdehyde (MDA), and 8-isoprostane levels in EBC | 25 Male COPD smokers | 26 Smokers and 29 non-smokers, males, healthy | EBC collected on an EcoScreen device, 15 min with nose clip, and then quickly stored at −70 °C for 6 months, MDA was measured using a commercial kit in a fluorescence detector in HPLC | Even if respiratory function tests were within normal limits, oxidant burden in the lungs of smokers was equivalent to that in COPD patients, 8-isoprostane could be useful in assessing symptom severity and health status of COPD patients | 16 Moderate |
| Larstad, 2001, Sweden [47] | To develop a method of MDA quantification | 29 Patients with asthma, 7 of which with wheezing | 15 control subjects without asthma | EBC was collected on an ECOSCREEN d evice, during 4 min, with a nose clip, stored at −20 °C until analysis, MDA was measured using TBS at 95 °C, and then separated by HPLC and MDA-TBA measured by fluorescence | No statistically significant difference between patients with asthma and patients without asthma; however, among females, subjects with asthma had higher MDA levels as compared with females without asthma; the use of the method when studying airway inflammation has to be further evaluated | 12 Moderate |
| Laumbach, 2014, USA [48] | To determine if exposure to traffic-related pollutant particles (TRAPs) during commuting causes acute oxidative stress in the respiratory tract or changes in heart rate variability | 21 Young volunteers (15 males and 6 females) | EBC was collected with an ECOSCREEN device, for 20 min, samples were frozen to −80 °C, MDA analysiswas performed by using a mixture of EBC, phosphoric acid, and thiobarbituric acid, which was heated and injected into an HPLC fluorescence system | Increases in markers of oxidative stress in EBC may represent early biological responses to widespread exposures to TRAPs particles that affect passengers in vehicles on heavily trafficked roadways | 9 Low | |
| Lee, 2015, Korea [49] | To investigate the actual health effects of multi wall carbon nanotubes in manufacturing workers | 9 Male carbon nanotube manufacturing workers, 5 smokers | 4 Office workers (3 males and 1 female), 2 smokers | EBC was collected on an Rtube, during 10 min, with a noseclip, stored in dry ice for 2 weeks. MDA was measured using fluorescence HPLC | Analyzed biomarkers in the MWCNT manufacturing workers were significantly higher than those in the office workers; MDA, n-hexanal, and molybdenum could be useful biomarkers of MWCNT exposure | 10 Moderate |
| Majewska, 2004, Poland [50] | To determine whether concentrations of H2O2 and TBARs in EBC are elevated and correlate with systemic response to pneumonia during 10 days of hospital treatment | 43 Inpatients with community acquired pneumonia (12 females and 31 males) | 20 Healthy non-smoking control subjects (6 females and 14 males) | EBC was collected on a device from Jaeger, cooled with ethanol at −9 °C, during 20 min, wearing a nose clip, stored on ice until measurement was performed, MDA was measured as TBARS | Pneumonia is accompanied by oxidative stress in airways that moderately correlates with intensity of systemic inflammatory response, determination of H2O2 in EBC may be helpful for noninvasive monitoring of oxidant production during lower respiratory tract infection | 13 Moderate |
| Nowak, 2001, Poland [51] | To investigate the concentration of H2O2 and TBARs in EBC and influencing factors | 58 Healthy volunteers (18 smokers and 40 non-smokers, 31 males and 27 females) | EBC was collected on a device from Jaeger Toennies, during 20 min, wearing a noseclip, MDA was measured as TBARS spectrofotometrically | Neither moderate exercise nor one puff of salbutamol nor ipratropium significantly influenced the concentration of H2O2 and TBARs in EBC, only 4 of 120 EBC specimens from non-smoker subjects revealed detectable levels of TBARs, cigarette smokers exhaled more TBARs | 11 Moderate | |
| Pelclova, 2018, Czech Republic [52] | To investigate lung oxidative stress in workers handling nanocomposites | 19 Nanocomposite-synthesizing and processing researchers (14 males and 5 females), all non-smokers | 19 Control subjects (13 males and 6 females, all non-smokers) | EBC was collected on an ECOSCREEN device, until a minimum of 120 L of exhaled breath, wearing a nose clip, immediately stored at −80 °C, MDA was measured | Significant associations were found between working in nanocomposite synthesis and EBC biomarkers; more research is needed to understand the contribution of nanoparticles from nanocomposite processing in inducing oxidative stress, relative to other co-exposures generated during welding, smelting, and secondary oxidation processes | 12 Moderate |
| Pelclova, 2014, Czech Republic [53] | To search for optimal markers in the exhaled breath condensate (EBC), plasma, and urine that would reflect the activity/severity of occupational asthma (OA) after withdrawal from the exposure to the allergen | 43 Subjects with previously diagnosed immunological OA (18 males and 25 females) | 20 Control subjects, working as office or healthcare employees and having no symptoms of asthma (10 males and 10 females) | EBC was collected for 15–20 min with EcoScreen, wearing a nose clip, samples were stored at −80 °C for a maximum of 2 months, MDA was measured with LC-ESI-MS/MS | Improvement in OA is very slow and objective impairments persist years after removal from the exposure; cysteinyl LTs and 8-ISO in EBC and 8-ISO in plasma might enrich the spectrum of useful objective tests for the follow-up of OA | 12 Moderate |
| Pelclova, 2016, Czech Republic [54] | To investigate the utility of oxidative stress biomarkers in EBC in iron oxide pigment production workers | 14 male workers, 43 ± 7 years, 43% smokers | 14 Males, 39 ± 4 years, 50% smokers, non-exposed | EBC was collected for 15 min with an EcoScreen device, wearing a nose clip, samples were stored at −80 °C, MDA was measured with LC-ESI-MS/MS | Almost all markers of lipid, nucleic acid, and protein oxidation were elevated in the EBC of workers as compared with control subjects; markers in urine were not elevated | 10 Moderate |
| Ricelli, 2018, Italy [55] | To investigate the cromium and nickel content of EBC of stainless steel (SS) tungsten inert gas (TIG) welders, and relate their concentrations with oxidative stress and inflammatory biomarkers | 100 SS welding workers, aged 18–65 years, smokers/non-smokers/ex-smokers 33/36/31, values were considered pre-shift | EBC was collected on an ECOSCREEN device, during 15 min, and stored at −80 °C, MDA was determined by tandem LC-MS/MS | Given the weak relationship between the biomarkers and effects of exposure, we speculate that other substances generated during SS TIG welding also play a role in generating lung oxidative stress | 11 Moderate | |
| Rundell, 2008, USA [56] | To investigate PM1 inhalation during exercise on lung function, exhaled nitric oxide (eNO), total nitrate (NO3), S-nitrosoglutathione (GSNO), and malondialdehyde (MDA) in EBC | Twelve physically fit, non-asthmatic, non-smoking males | EBC was collected on an ECOSCREEN device, during 15 min, wearing a nose clip and stored at −80 °C, MDA was measured after derivatization with TBA by HPLC fluorescence | MDA increased 40% after low PM exercise, high PM1 inhalation during exercise caused a reduced alveolar contribution to eNO, NO3 and eNO variables were decreased and were related to impaired lung function | 9 Low | |
| Sakhvidi, 2015, Iran [57] | To measure exhaled breath malondialdehyde (EBC-MDA) in workers exposed to dust containing silica as compared with that of a non-exposed control group | 25 Male workers in a ceramic factory | 25 Male control subjects from administrative departments of the same factory | EBC was collected with a homemade apparatus, during 5 min, wearing a nose clip, samples were stored in a freezer at −20 °C until examination, MDA was analyzed using an HPLC equipped with a fluorescent detector | Significant correlation between respirable dust exposure intensity and the level of EBC-MDA of the exposed subjects, but no significant correlation with lung functions | 13 Moderate |
| Sauvain, 2014, Switzerland [58] | To evaluate the feasibility of using exhaled breath condensate (EBC) from healthy volunteers for (1) assessing the lung deposited dose of combustion nanoparticles and (2) determining the resulting oxidative stress | 15 Healthy non-smoking control subjects | EBC collected on a Rtube, during 10 min, wearing a nose clip, a aliquote was stored at −78 °C, MDA was derivatized with TBA, and then measured with HPLC fluorescence | The results suggest two phases of oxidation markers in EBC: first, the initial deposition of particles and gases in the lung lining liquid, and later the start of oxidative stress with associated cell membrane damage | 11 Moderate | |
| Stockfelt, 2012, Sweden [59] | To examine airway effects of two kinds of wood smoke in a chamber study | 16 Healthy non-smoking control subjects (8 males and 8 females), 3 excluded for respiratory problems before the experiment | EBC collected according to Barregard, MDA measured always according to Barregard | Relatively low levels of wood smoke exposure induce effects on airways | 8 Low | |
| Syslova, 2009, Czech Republic [60] | To develop a sensitive method for a parallel, rapid and precise determination of the most prominent oxidative stress biomarkers for patients with silica or asbestos disease | 20 Subjects with previous exposure to silica or asbestos and related disease | 10 Control subjects | EBC was collected on an ECOSCREEN device, for 5−10 min, wearing a nose clip, frozen at −80 °C, and stored for a maximum of 1 month, MDA was measured with optimized LC–ESI-MS/MS | The differences in concentration levels of biomarkers between the two groups was perceptible in all the body fluids (the difference observed in an exhaled breath condensatewas statistically most significant) | 12 Moderate |
| Szkudlarek, 2003, Poland [61] | To test whether exhalation of H2O2 and TBARs by healthy subjects depends on reactive oxygen species generation from blood phagocytes | 41 Healthy, non-smoking control subjects, mean age 20.770.8 years (18 males and 23 females) | EBC was collected on a homemade mouth piece connected to a glass tube cooler, wearing a nose clip, during 20 min, MDA was measured as TBARS according to Nowak | No association between exhaled TBARs and blood phagocytes activity was found | 10 Moderate | |
| Pelclova, 2020, Czech Republic [62] | To analyze biomarkers in EBC of nanocomposite workers and understand the health effects | 20 Researchers handling nanocomposites (15 males and 5 females), 19 non-smokers, mean age of 41.8 years | 21 Control subjects (15 males and 6 females), office emplyees, 19 non-smokers, mean age of 42.7 years | EBC was collected on ECOSCREEN and stored at −80 °C, MDA was measured with LC-ESI- MS/MS | among inflammation markers, LT4 and tumor necrosis factor were the most useful | 10 moderate |