Defining adult asthma endotypes by clinical features and patterns of volatile organic compounds in exhaled air

Background

Several classifications of adult asthma patients using cluster analyses based on clinical and demographic information has resulted in clinical phenotypic clusters that do not address molecular mechanisms. Volatile organic compounds (VOC) in exhaled air are released during inflammation in response to oxidative stress as a result of activated leukocytes. VOC profiles in exhaled air could distinguish between asthma patients and healthy subjects. In this study, we aimed to classify new asthma endotypes by combining inflammatory mechanisms investigated by VOC profiles in exhaled air and clinical information of asthma patients.

Methods

Breath samples were analyzed for VOC profiles by gas chromatography–mass spectrometry from asthma patients (n = 195) and healthy controls (n = 40). A total of 945 determined compounds were subjected to discriminant analysis to find those that could discriminate healthy from asthmatic subjects. 2-step cluster analysis based on clinical information and VOCs in exhaled air were used to form asthma endotypes.

Results

We identified 16 VOCs, which could distinguish between healthy and asthma subjects with a sensitivity of 100% and a specificity of 91.1%. Cluster analysis based on VOCs in exhaled air and the clinical parameters FEV1, FEV1 change after 3 weeks of hospitalization, allergic sensitization, Junipers symptoms score and asthma medications resulted in the formation of 7 different asthma endotype clusters. We identified asthma clusters with different VOC profiles but similar clinical characteristics and endotypes with similar VOC profiles, but distinct clinical characteristics.

Conclusion

This study demonstrates that both, clinical presentation of asthma and inflammatory mechanisms in the airways should be considered for classification of asthma subtypes.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-014-0136-8) contains supplementary material, which is available to authorized users.     

Measurement of volatile organic compounds in exhaled breath as collected in evacuated electropolished canisters

A set of three complementary analytical methods were developed specifically for exhaled breath as collected in evacuated stainless steel canisters using gas chromatographic-mass spectrometric detection. The first is a screening method to quantify the carbon dioxide component (generally at 4–5% concentration), the second method measures the very volatile high-level endogenous compounds [e.g. acetone and isoprene at 500–1000 parts per billion by volume (ppbv), methanol, ethanol, dimethylsulfide at 2–10 ppbv], and the third method is designed to measure trace-level environmental contaminants and other endogenous volatile organic compounds (VOCs) (sub-ppbv) in breath. The canister-based sample format allows all three methods to be applied to each individual sample for complete constituent characterization. Application of these methods is shown to be useful in the following ways: analysis of CO₂ levels indicates the approximate quantity of alveolar breath collected (as opposed to whole breath) in a sample; levels of major endogenous compounds are shown to be influenced by physical activities and subsequent recovery periods; and environmental exposures to xenobiotic VOCs can be characterized by assessment of post-exposure breath elimination curves. The instrumentation and methodology are described and example chromatograms and quantitative data plots demonstrating the utility of the methods are presented.     

Biofiltration of volatile organic compounds

The removal of volatile organic compounds (VOCs) from contaminated airstreams has become a major air pollution concern. Improvement of the biofiltration process commonly used for the removal of odorous compounds has led to a better control of key parameters, enabling the application of biofiltration to be extended also to the removal of VOCs. Moreover, biofiltration, which is based on the ability of micro-organisms to degrade a large variety of compounds, proves to be economical and environmentally viable. In a biofilter, the waste gas is forced to rise through a layer of packed porous material. Thus, pollutants contained in the gaseous effluent are oxidised or converted into biomass by the action of microorganisms previously fixed on the packing material. The biofiltration process is then based on two principal phenomena: (1) transfer of contaminants from the air to the water phase or support medium, (2) bioconversion of pollutants to biomass, metabolic end-products, or carbon dioxide and water. The diversity of biofiltration mechanisms and their interaction with the microflora mean that the biofilter is defined as a complex and structured ecosystem. As a result, in addition to operating conditions, research into the microbial ecology of biofilters is required in order better to optimise the management of such biological treatment systems.     

Preliminary investigation of using volatile organic compounds from human expired air, blood and urine for locating entrapped people in earthquakes

A preliminary investigation on the possibility of using volatile organic compounds (VOCs) determination of expired air, blood and urine, for the early location of entrapped people in earthquakes, has been carried out. A group of 15 healthy subjects has been sampled. The identification of a common "core" of substances might provide indications of human presence that can be used for the development of a real time field analytical method for the on site detection of entrapped people. Expired air samples have been analyzed by thermal desorption GC/MS and VOCs from blood and urine by headspace SPME-GC/MS. Acetone was the only compound found common in all three matrices. Isoprene was found in both expired air and blood samples. Acetone and isoprene along with a number of saturated hydrocarbons were among the major constituents identified in expired air analysis. Various ketones (2-pentanone, 4-heptanone, 2-butanone) were also determined over urine specimens. Using the techniques and methods of field analytical chemistry and technology appears to be the proper approach for applying the results of the present study in real situations.     

The volatile organic compounds from the mycelium of Tuber borchii Vitt

The mycelium of T. borchii (characterized by DNA analysis) grown in sterile liquid medium produced some VOCs. The VOCs were retained on carbographs by passing a flow of helium, isolated and characterized in a GC-MS equipment after a thermal desorption. The compounds present in the VOCs from the mycelium cultures, but not in the VOCs from the control cultures, contained 29 compounds. The main compounds were 1,3-ditertbutylbenzene (16.1 ng/l), 3-methylheptane (9.2 ng/l), butan-2-one (8.8 ng/l), ethynylbenzene (5.6 ng/l), and octan-3-one (4.9 ng/l).     

A novel method for characterisation of microbial growth kinetics on volatile organic compounds

A novel method for the determination of microbial growth kinetics on hydrophobic volatile organic compounds (VOC) has been developed. A stirred tank reactor was operated as a fed-batch system to which the VOC was continuously fed via the gas phase, assuring a constant VOC concentration in the mineral medium. A flow of air was saturated with the VOC, and then mixed with a further flow of air, to obtain a predetermined VOC concentration. Thus, different VOC concentrations in the mineral medium could be obtained by altering the VOC concentration in the feed gas. The growth kinetics of Xanthobacter autotrophicus GJ10 on 1,2-dichloroethane (DCE) and of Pseudomonas sp. strain JS150 on MonoChloroBenzene (MCB) were assessed using this method. The growth of strain JS150 was strongly inhibited at MCB concentrations higher than 160 mg l⁻¹, and the results were fitted using a piecewise function. The growth kinetics of strain GJ10 were described by the Luong model where maximum growth rate μ_max = 0.12 h⁻¹, substrate saturation constant K _S = 7.8 mg l⁻¹, and maximum substrate concentration S _m (above which growth is completely inhibited) = 1080 mg l⁻¹. Varying nitrogen and oxygen flows enabled the effect of oxygen concentration on the growth kinetics of Pseudomonas JS150 to be determined. Received: 30 November 1998 / Received revision: 19 March 1999 / Accepted: 20 March 1999     

A review of the USEPA's single breath canister (SBC) method for exhaled volatile organic biomarkers

Exhaled alveolar breath can provide a great deal of information about an individual's health and previous exposure to potentially harmful xenobiotic materials. Because breath can be obtained non-invasively and its constituents directly reflect concentrations in the blood, its use has many potential applications in the field of biomarker research. This paper reviews the utility and application of the single breath canister (SBC) method of alveolar breath collection and analysis first developed by the US Environmental Protection Agency (USEPA) in the 1990s. This review covers the development of the SBC technique in the laboratory and its application in a range of field studies. Together these studies specifically show how the SBC method (and exhaled breath analysis in general) can be used to clearly demonstrate recent exposure to volatile organic compounds, to link particular activities to specific exposures, to determine compoundspecific uptake and elimination kinetics, and to assess the relative importance of various routes of exposure (i.e. dermal, ingestion, inhalation) in multipathway scenarios. Specific investigations covered in this overview include an assessment of exposures related to the residential use of contaminated groundwater, exposures to gasoline and fuel additives at self-service gas stations, swimmers' exposures to trihalomethanes, and occupational exposures to jet fuel.     

A review of the USEPA's single breath canister (SBC) method for exhaled volatile organic biomarkers

Exhaled alveolar breath can provide a great deal of information about an individual's health and previous exposure to potentially harmful xenobiotic materials. Because breath can be obtained non-invasively and its constituents directly reflect concentrations in the blood, its use has many potential applications in the field of biomarker research. This paper reviews the utility and application of the single breath canister (SBC) method of alveolar breath collection and analysis first developed by the US Environmental Protection Agency (USEPA) in the 1990s. This review covers the development of the SBC technique in the laboratory and its application in a range of field studies. Together these studies specifically show how the SBC method (and exhaled breath analysis in general) can be used to clearly demonstrate recent exposure to volatile organic compounds, to link particular activities to specific exposures, to determine compoundspecific uptake and elimination kinetics, and to assess the relative importance of various routes of exposure (i.e. dermal, ingestion, inhalation) in multipathway scenarios. Specific investigations covered in this overview include an assessment of exposures related to the residential use of contaminated groundwater, exposures to gasoline and fuel additives at self-service gas stations, swimmers' exposures to trihalomethanes, and occupational exposures to jet fuel.     

Abatement of volatile organic sulfur compounds in odorous emissions from the bio-industry

Compounds of interest in this work are methanethiol (MeSH), dimethyl sulfide (Me₂S), dimethyl polysulfides (Me₂S_x) and carbon disulfide (CS₂) since these volatiles have been identified as predominant odorants in the emission of a wide range of activities in the bio-industry (e.g. aerobic waste water treatment plants, composting plants, rendering plants). In these processes, the occurrence of volatile organic sulfur compounds is mainly related to the presence of anaerobic microsites with consecutive fermentation of sulfur containing organic material and/or to the breakdown of the latter due to thermal heating. Due to the chemical complexity of these low-concentrated waste gas streams and the high flow rates to be handled, mainly biotechnological techniques and scrubbers can be used to control the odour emission. When using biofilters or trickling filters, inoculation with specific micro-organisms and pH-control strategies should be implemented to optimise the removal of volatile organic sulfur compounds. In scrubbers, chemical oxidation of the volatile organic sulfur compounds can be obtained by dosing hypochlorite, ozone or hydrogen peroxide to the scrubbing liquid. However, optimal operational conditions for each of these abatement techniques requires a further research in order to guarantee a long-term and efficient overall odour abatement.     

基于硅胶管吸附萃取的热脱附-气相色谱-质谱联用法测定真菌的挥发性有机化合物

【背景】真菌代谢产生的挥发性有机化合物(Volatile organic compounds,VOCs)具有重要的生物学功能。高通量的VOCs测定方法对于VOCs产生菌的筛选非常重要。【目的】建立一种简单可靠、基于硅胶管吸附萃取(Siliconetubesorptiveextraction,STSE)的热脱附-气相色谱-质谱(Thermal desorption-gas chromatography-mass spectrometry,TD-GC-MS)联用法测定真菌培养物VOCs的高通量分析技术。【方法】选择代表性的包括酯、酮、醇、醛、烯等21种VOCs作为对照品建立标准曲线;将对照品滴加于PDA培养基,基于STSE进行TD-GC-MS分析,测定其专属性;将木霉菌T552接种于PDA培养基,基于STSE进行TD-GC-MS分析,测定方法的重复性;在以上基础上,用上述试验条件对6株木霉菌产生的VOCs进行筛选。【结果】在全扫描和提取离子条件下,21种对照品分别在1.0-10.0μg/mL、0.1-10.0μg/mL范围内线性关系良好。基于STSE的TD-GC-MS分析发现,滴加在PDA培养基上的对照品分离良好,且PDA培养基平板中的其它物质不干扰VOCs的测定,表明该方法专属性良好;对PDA培养的木霉菌T552进行的VOCs重复测定鉴定的5种VOCs相对标准偏差均小于10%,表明该方法重复性良好。用建立的基于STSE的TD-GC-MS方法从6株供试木霉菌中鉴定出包括酸、醇、酯、烯等共37种VOCs。【结论】建立了一种简单可靠、基于STSE的TD-GC-MS分析测定真菌培养物VOCs的高通量分析技术,可用于真菌VOCs的分析测定。     

Production of volatile organic compounds by mycobacteria

The need for improved rapid diagnostic tests for tuberculosis disease has prompted interest in the volatile organic compounds (VOCs) emitted by Mycobacterium tuberculosis complex bacteria. We have investigated VOCs emitted by Mycobacterium bovis BCG grown on Lowenstein-Jensen media using selected ion flow tube mass spectrometry and thermal desorption-gas chromatography-mass spectrometry. Compounds observed included dimethyl sulphide, 3-methyl-1-butanol, 2-methyl-1-propanol, butanone, 2-methyl-1-butanol, methyl 2-methylbutanoate, 2-phenylethanol and hydrogen sulphide. Changes in levels of acetaldehyde, methanol and ammonia were also observed. The compounds identified are not unique to M.?bovis BCG, and further studies are needed to validate their diagnostic value. Investigations using an ultra-rapid gas chromatograph with a surface acoustic wave sensor (zNose) demonstrated the presence of 2-phenylethanol (PEA) in the headspace of cultures of M.?bovis BCG and Mycobacterium smegmatis, when grown on Lowenstein-Jensen supplemented with glycerol. PEA is a reversible inhibitor of DNA synthesis. It is used during selective isolation of gram-positive bacteria and may also be used to inhibit mycobacterial growth. PEA production was observed to be dependent on growth of mycobacteria. Further study is required to elucidate the metabolic pathways involved and assess whether this compound is produced during in vivo growth of mycobacteria.     

Elimination of volatile compounds of leaf tobacco from air emissions using biofiltration

The composition of the volatile organic compounds (VOCs) of various leaf tobacco brands and their blends has been studied. The differences in the content of nicotine, solanone, tetramethyl hexadecenol, megastigmatrienones, and other compounds, determining the specific tobacco smell, have been revealed. A microbial consortium, which is able to deodorize simulated tobacco emissions and decompose nicotine, has been formed by long-term adaptation to the VOCs of tobacco leaves in a laboratory reactor, functioning as a trickle-bed biofilter. Such a biofilter eliminates 90% of the basic toxic compound (nicotine) and odor-active compounds; the filtration efficiency does not change for tobacco brands with different VOC concentrations or in the presence of foreign substances. The main strains, isolated from the formed consortium and participating in the nicotine decomposition process, belong to the genera Pseudomonas, Bacillus, and Rhodococcus. An examination of the biofilter trickling fluid has shown full decomposition of nicotine and odor-active VOCs. The compounds, revealed in the trickling fluid, did not have any odor and were nontoxic. The obtained results make it possible to conduct scaling of the biofiltration process to eliminate odor from air emissions in the tobacco industry.     

Identification of multiple symptoms of huanglongbing by electronic nose based on the variability of volatile organic compounds

Huanglongbing (HLB) is highly contagious and cannot be cured, resulting in a decrease in the commercial value of citrus. Timely detection and removal of diseased trees is an effective way to reduce losses. Complex symptoms of HLB, such as nutrient deficiencies often accompany HLB; as a result effective and accurate identification of HLB remains a challenge. In this study, 175 volatile organic compounds (VOCs) were detected in three categories (healthy, HLB, and Zn-deficiency) of samples using headspace solid-phase microextraction gas chromatography–mass spectrometry (HS-SPME-GC/MS), highlighting the variability of VOCs present in different categories of samples. In order to simplify the testing steps and reduce the cost in practical agricultural production, a method based on electronic nose technology to collect VOCs from citrus leaves for HLB detection was proposed. Among them, limiting value features and linear discriminant analysis were identified as the best combination of feature extraction and pattern recognition methods. Multiple sets of comparison experiments were set up and the collection conditions of VOCs were optimized. The results showed that the best classification performance was achieved for a 0.2 g sample at a collection time of 20 min when the collection temperature was 40°C and the headspace volume was 200 mL. Four types of samples (healthy, HLB-positive, Zn-deficiency, Zn-deficiency and HLB-positive) were used for model reliability validation, with an accuracy of 97.79% for HLB samples for multiple symptoms (including HLB-positive and Zn-deficiency and HLB-positive) identification. In addition, the accuracy of samples with a combined effect of Zn-deficiency and HLB was 96.43%. The results show that the E-nose-based HLB detection method is conducive to suppressing the spread of HLB, which can ensure the quality of citrus products and reduce the economic loss to horticulturists, and has good practical value.     

Detection of volatile organic compounds released by wood furniture based on a cataluminescence test system

Wood furniture is an important source of indoor air pollution. To date, the detection of harmful substances in wood furniture has relied on the control of a single formaldehyde component, therefore the detection and evaluation of pollutants released by wood furniture are necessary. A novel method based on a cataluminescence (CTL) sensor system generated on the surface of nano‐3TiO₂–2BiVO₄ was proposed for the simultaneous detection of pollutants released by wood furniture. Formaldehyde and benzene were selected as a model to investigate the CTL‐sensing properties of the sensor system. Field emission scanning electronic microscopy (FESEM), transmission electron microscopy (TEM) and X‐ray diffraction (XRD) were employed to characterize the as‐prepared samples. The results showed that the as‐prepared test system exhibited outstanding CTL properties such as stable intensity, a high signal‐to‐noise ratio, and short response and recovery times. In addition, the limit of detection for formaldehyde and benzene was below the standard permitted concentrations. Moreover, the sensor system showed outstanding selectivity for formaldehyde and benzene compared with eight other common volatile organic compounds (VOCs). The performance of the sensor system will enable furniture VOC limit emissions standards to be promulgated as soon as possible. Copyright © 2015 John Wiley & Sons, Ltd.     

Detection and analysis of endogenous polar volatile organic compounds (PVOCs) in urine for human exposome research

Background: The human exposome, defined as ‘…everything that is not the genome’, comprises all chemicals in the body interacting with life processes. The exposome drives genes x environment (GxE) interactions that can cause long-term latency and chronic diseases. The exposome constantly changes in response to external exposures and internal metabolism. Different types of compounds are found in different biological media.

Objective: Measure polar volatile organic compounds (PVOCs) excreted in urine to document endogenous metabolites and exogenous compounds from environmental exposures.

Methods: Use headspace collection and sorbent tube thermal desorption coupled with bench-top gas chromatography-mass spectrometry (GC-MS) for targeted and non-targeted approaches. Identify and categorize PVOCs that may distinguish among healthy and affected individuals.

Results: Method is successfully demonstrated to tabulate a series of 28 PVOCs detected in human urine across 120 samples from 28 human subjects. Median concentrations range from below detect to 165?ng/mL. Certain PVOCs have potential health implications.

Conclusions: Headspace collection with sorbent tubes is an effective method for documenting PVOCs in urine that are otherwise difficult to measure. This methodology can provide probative information regarding biochemical processes and adverse outcome pathways (AOPs) for toxicity testing.