Non-invasive spatial visualization system of exhaled ethanol for real-time analysis of ALDH2 related alcohol metabolism

A novel imaging system of ethanol in exhaled breath induced by acetaldehyde dehydrogenase (ALDH2)-related alcohol metabolism has been developed. The system provides an image of ethanol distribution as chemiluminescence (CL) on an enzyme-immobilized support. The spatiotemporal change of CL generated by ethanol in exhaled breath after oral administration of ethanol was detected by employing an electron multiplier CCD (EM-CCD) camera, illustrated and analyzed. Prior to measurement of standard gaseous ethanol and ethanol in exhaled breath, the system was optimized by investigating the enzyme-immobilized supports, concentration of substrate and pH condition of Tris-HCl buffer solution. The ethanol skin patch test, a simple method as an indicator of ALDH2, was performed on healthy volunteers. Breath samples of 5 volunteers with ALDH2 (+) and 5 volunteers with ALDH2 (-) were used for exhaled ethanol analysis. Concentration-time profiles of exhaled ethanol obtained from all volunteers were analyzed over a period of 120 min after oral administration of ethanol (0.4 g per kg body weight) in the form of beer which contains 5% of alcohol. The results obtained from the system showed that the peaks of exhaled ethanol concentrations appeared at 30 min, which was considered as a rapid ethanol absorption phase following first-order kinetics. Exhaled ethanol concentrations of volunteers with ALDH2 (+) were lower than volunteers with ALDH2 (-) and the digestion of ethanol in volunteers with ALDH2 (+) was faster than in volunteers with ALDH2 (-). The eliminations were analyzed to follow zero-order kinetics with a rate constant for each group.     

An exploratory comparative study of volatile compounds in exhaled breath and emitted by skin using selected ion flow tube mass spectrometry

Selected ion flow tube mass spectrometry (SIFT-MS) has been used to carry out a pilot parallel study on five volunteers to determine changes occurring in several trace compounds present in exhaled breath and emitted from skin into a collection bag surrounding part of the arm, before and after ingesting 75 g of glucose in the fasting state. SIFT-MS enabled real-time quantification of ammonia, methanol, ethanol, propanol, formaldehyde, acetaldehyde, isoprene and acetone. Following glucose ingestion, blood glucose and trace compound levels were measured every 30 min for 2 h. All the above compounds, except formaldehyde, were detected at the expected levels in exhaled breath of all volunteers; all the above compounds, except isoprene, were detected in the collection bag. Ammonia, methanol and ethanol were present at lower levels in the bag than in the breath. The aldehydes were present at higher levels in the bag than in breath. The blood glucose increased to a peak about 1 h post-ingestion, but this change was not obviously correlated with temporal changes in any of the compounds in breath or emitted by skin, except for acetone. The decrease in breath acetone was closely mirrored by skin-emitted acetone in three volunteers. Breath and skin acetone also clearly change with blood glucose and further work may ultimately enable inferences to be drawn of the blood glucose concentration from skin or breath measurements in type 1 diabetes.     

A selected ion flow tube mass spectrometry study of ammonia in mouth- and nose-exhaled breath and in the oral cavity

A study has been carried out, involving three healthy volunteers, of the ammonia levels in breath exhaled via the mouth and via the nose and in the static oral cavity using on-line, selected ion flow tube mass spectrometry (SIFT-MS), obviating the problems associated with sample collection of ammonia. The unequivocal conclusion drawn is that the ammonia appearing in the mouth-exhaled breath of the three volunteers is largely generated in the oral cavity and that the ammonia originating at the alveolar interface in the lungs is typically at levels less than about 100 parts-per-billion, which is a small fraction of the total breath ammonia. This leads to the recommendation that exhaled breath analyses should focus on nose-exhaled breath if the objective is to use breath analysis to investigate systemic, metabolic disease.     

Noninvasive detection of colorectal cancer by analysis of exhaled breath

There has been growing interest in exhaled breath analysis for cancer screening and disease monitoring; however, limited breath biomarker information exists regarding colorectal cancer (CRC). The objective of this study was to screen for breath biomarkers of CRC. Exhaled breath was collected from 20 CRC patients and 20 healthy controls; subsequently, solid-phase microextraction–gas chromatography/mass spectrometry (SPME-GC/MS) was used to assess the exhaled volatile organic compounds (VOCs) of the study participants. The statistical methods of principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were performed to process the final data. The VOCs in the exhalations of CRC patients exhibited significant differences from the VOCs in the exhalations of healthy controls; in particular, relative to the latter exhalations, the former exhalations contain significantly higher levels of cyclohexanone, 2,2-dimethyldecane, dodecane, 4-ethyl-1-octyn-3-ol, ethylaniline, cyclooctylmethanol, trans-2-dodecen-1-ol, and 3-hydroxy-2,4,4-trimethylpentyl 2-methylpropanoate but significantly lower levels of 6-t-butyl-2,2,9,9-tetramethyl-3,5-decadien-7-yne (P?<?0.05). Analyses of breath VOCs provide a related model of CRC exhalation that could represent an effective and convenient screening method for this disease.

Selected ion flow tube mass spectrometry of exhaled breath condensate headspace

Collection of exhaled breath condensate (EBC) is a relatively simple noninvasive method of breath analysis; however, no data have been reported that would relate concentration of volatile compounds in EBC to their gaseous concentrations in exhaled air. The aim of the study was to investigate which volatile compounds are present in EBC and how their concentrations relate to results of direct breath analysis. Thus, samples of EBC were collected in a standard way from several subjects and absolute levels of several common volatile breath metabolites (ammonia, acetone, ethanol, methanol, propanol, isoprene, hydrogen cyanide, formaldehyde and acetaldehyde) were then determined in their headspace using selected ion flow tube mass spectrometry (SIFT-MS). Results are compared with those from on-line breath analyses carried out immediately before collecting the EBC samples. It has been demonstrated that SIFT-MS can be used to quantify the concentrations of volatiles in EBC samples and that, for methanol, ammonia, ethanol and acetone, the EBC concentrations correlate with the direct breath levels. However, the EBC concentrations of isoprene, formaldehyde, acetaldehyde, hydrogen cyanide and propanol do not correlate with direct breath measurements. Copyright (c) 2008 John Wiley & Sons, Ltd.     

A longitudinal study of ethanol and acetaldehyde in the exhaled breath of healthy volunteers using selected-ion flow-tube mass spectrometry

Selected-ion flow-tube mass spectrometry (SIFT-MS) has been used to monitor the volatile compounds in the exhaled breath of 30 volunteers (19 male, 11 female) over a 6-month period. Volunteers provided breath samples each week between 8:45 and 13:00 (before lunch), and the concentrations of several trace compounds were obtained. In this paper the focus is on ethanol and acetaldehyde, which were simultaneously quantified by SIFT-MS using H3O+ precursor ions. The mean ethanol level for all samples was 196 parts-per-billion (ppb) with a standard deviation of 244 ppb, and the range of values for breath samples analysed is 0 to 1663 ppb. The mean acetaldehyde level for all samples was 24 ppb with a standard deviation of 17 ppb, and the range of values for breath samples analysed is 0 to 104 ppb. Background (ambient air) levels of ethanol were around 50 ppb, whereas any background acetaldehyde was usually undetectable. Increased ethanol levels were observed if sweet drink/food had been consumed within the 2 h prior to providing the breath samples, but no increase was apparent when alcohol had been consumed the previous evening. The measured endogenous breath ethanol and acetaldehyde levels were not correlated. These data relating to healthy individuals are a prelude to using breath analysis for clinical diagnosis, for example, the recognition of bacterial overload in the gut (ethanol) or the possibly of detecting tumours in the body (acetaldehyde).     

Detection of acetone in the human breath as diabetes biomarker based on PPy/WO3 nanocomposite sensor by FFT continuous cyclic voltammetry method

This research represents a novel detection method of acetone level in the exhaled breath samples (RH=88 %) based on polypyrrole/tungsten oxide (PPy/WO₃) nanocomposite sensor. The PPy/WO₃ sensor was fabricated by the deposition of nanocomposite on/between interdigitated electrodes (IDEs) through electrospray coating and was then characterized by FE-SEM imaging. In this detection method, the coulometric signal of the sensor was calculated using Fast Fourier Continuous Cyclic Voltammetry (FFTCCV), where cyclic voltammetry (CV) was applied to the sensor in the defined potential rang and then charge changes of the sensor was obtained by integration of the current in all scanned potential ranges. FFTCCV method enhances the sensitivity of the sensor when exposed to the gas mixtures containing acetone. In addition to its fast coulometric response time (≤5 s) in the two linear ranges of 0.7–2.8 ppm and 2.8–28.2 ppm (R²=0.99), FFTCCV method provides the low detection limit of 70 ppb, and high sensitivity toward acetone at the optimum values of the parameters. The fabricated sensor showed great selectivity toward acetone when exposed to humid air and some exhaled gas like carbon dioxide, ammonia, methanol, ethanol and alkyl amines. The results were very satisfying as the sensor was capable to detect different acetone levels in human exhaled breath as non-invasive diagnosis of diabetes with a good correlation (R²≃0.9) to the routine blood sugar test taken by different commercial glucometers results.     

Investigation of acetone,butanol and carbon dioxide as new breath biomarkers for convenient and noninvasive diagnosis of obstructive sleep apnea syndrome

The objective of the present study was to investigate whether analysis of carbon dioxide, acetone and/or butanol present in human breath can be used as a simple and noninvasive diagnosis method for obstructive sleep apnea syndrome (OSAS). For this purpose, overnight changes in the concentrations of these breath molecules were measured before and after sleep in 10 patients who underwent polysomnography and were diagnosed with OSAS, and were compared with the levels of these biomarkers determined after sleep in 10 healthy subjects. The concentrations of exhaled carbon dioxide were measured using external cavity laser‐based off‐axis cavity enhanced absorption spectroscopy, whereas the levels of exhaled acetone and butanol were determined using thermal desorption gas chromatography mass spectrometry. We observed no significant changes in the levels of exhaled acetone and carbon dioxide in OSAS patients after sleep compared with pre‐sleep values and compared with those in healthy control subjects. However, for the first time, to our knowledge, analyses of expired air showed an increased concentration of butanol after sleep compared with that before sleep and compared with that in healthy subjects. These results suggest that butanol can be established as a potential biomarker to enable the convenient and noninvasive diagnosis of OSAS in the future.     

Performance of Amperometric Platinized‐Nafion Based Gas Phase Sensor for Determining Nitric Oxide (NO) Levels in Exhaled Human Nasal Breath

Nitric oxide (NO) levels in exhaled breath are a non‐invasive marker that can be used to diagnose various respiratory diseases and monitor a patient's response to given therapies. A portable and inexpensive device that can enable selective NO concentration measurements in exhaled breath samples is needed. Herein, the performance of an amperometric Pt‐Nafion‐based gas phase sensor for detection of NO in exhaled human nasal breath is examined. Enhanced selectivity over carbon monoxide and ammonia is achieved via an in‐line zinc oxide‐based filter. Exhaled nasal NO levels measured in 21 human samples with the sensor are shown to correlate well with those obtained using a chemiluminescence reference method (R²=0.9836).     

Direct and Selective Production of Propene from Bio‐Ethanol on Sc‐Loaded In2O3 Catalysts

Propene, one of key building blocks for manufacturing plastics and chemicals, could be directly and stably produced from ethanol in good yields. The conversion degree of ethanol to propene reached approximately 60 mol % by using a 3 atom % scandium‐loaded indium oxide catalyst at 823 K in the presence of water and hydrogen. The introduction of Sc prevented the reduction of In₂O₃ to In metal during the reaction, and that of water decreased the coke formation. Both additions resulted in longer lifetimes of the catalysts. The hydrogen addition increased the conversion of acetone to propene. The reaction pathways are also suggested on the basis of the product distributions and the pulse experiments, ethanol→acetaldehyde→acetone→propene, which is quite different from the shape‐selective catalysis on zeolites and the dimerization‐metathesis of ethene on nickel ion‐loaded silica catalysts.     

Ultrafast gas chromatography coupled to electronic nose to identify volatile biomarkers in exhaled breath from chronic obstructive pulmonary disease patients: A pilot study

An analytical method to identify volatile organic compounds (VOCs) in the exhaled breath from patients with a diagnosis of chronic obstructive pulmonary disease (COPD) using a ultrafast gas chromatography system equipped with an electronic nose detector (FGC eNose) has been developed. A prospective study was performed in 23 COPD patients and 33 healthy volunteers; exhalation breathing tests were performed with Tedlar bags. Each sample was analyzed by FCG eNose and the identification of VOCs was based on the Kovats index. Raw data were reduced by principal component analysis (PCA) and canonical discriminant analysis [canonical analysis of principal coordinates (CAP)]. The FCG eNose technology was able to identify 17 VOCs that distinguish COPD patients from healthy volunteers. At all stages of PCA and CAP the discrimination between groups was obvious. Chemical prints were correctly classified up to 82.2%, and were matched with 78.9% of the VOCs detected in the exhaled breath samples. Receiver operating characteristic curve analysis indicated the sensitivity and specificity to be 96% and 91%, respectively. This pilot study demonstrates that FGC eNose is a useful tool to identify VOCs as biomarkers in exhaled breath from COPD patients. Further studies should be performed to enhance the clinical relevance of this quick and ease methodology for COPD diagnosis.     

Chromatographic analysis of VOC patterns in exhaled breath from smokers and nonsmokers

Cigarette smoking harms nearly every organ of the body and causes many diseases. The analysis of exhaled breath for exogenous and endogenous volatile organic compounds (VOCs) can provide fundamental information on active smoking and insight into the health damage that smoke is creating. Various exhaled VOCs have been reported as typical of smoking habit and recent tobacco consumption, but to date, no eligible biomarkers have been identified. Aiming to identify such potential biomarkers, in this pilot study we analyzed the chemical patterns of exhaled breath from 26 volunteers divided into groups of nonsmokers and subgroups of smokers sampled at different periods of withdrawal from smoking. Solid‐phase microextraction technique and gas chromatography/mass spectrometry methods were applied. Many breath VOCs were identified and quantified in very low concentrations (ppbv range), but only a few (toluene, pyridine, pyrrole, benzene, 2‐butanone, 2‐pentanone and 1‐methyldecyclamine) were found to be statistically significant variables by Mann–Whitney test. In our analysis, we did not consider the predictive power of individual VOCs, as well as the criterion of uniqueness for biomarkers suggests, but we used the patterns of the only statistically significant compounds. Probit prediction model based on statistical relevant VOCs‐patterns showed that assessment of smoking status is heavily time dependent. In a two‐class classifier model, it is possible to predict with high specificity and sensitivity if a subject is a smoker who respected 1 hour of abstinence from smoking (short‐term exposure to tobacco) or a smoker (labelled "blank smoker") after a night out of smoking (long‐term exposure to tobacco). On the other side, in our study "blank smokers" are more like non‐smokers so that the two classes cannot be well distinguished and the corresponding prediction results showed a good sensitivity but low selectivity.     

Solvent effect on the alkaline hydrolysis of 2‐thiophenyl‐ 3,5‐dinitropyridine

The kinetics of the alkaline hydrolysis of 2‐thiophenyl‐3,5‐dinitropyridine were studied spectrophotometrically in different aquo‐organic solvents such as methanol, ethanol, n‐propyl alcohol, iso‐propyl alcohol, t‐butyl alcohol, acetonitrile, dimethyl sulfoxide, dioxane, and acetone at 30°C with various solvent compositions up to 80% (v/v) of organic components. An increase in the organic solvent percentage (v/v) has different effects on the reaction rate constants presumably due to hydrogen bond donor HBD and acceptor HBA of the medium and other solvatochromic parameters. Linear and nonlinear plots of log k against the reciprocal of the dielectric constant of the solvent were obtained. The effects are too complex to be analyzed in terms of a single parameter, but an approach using the Kamlet–Taft solvatochromic parameters is applied successfully to six mixed aquo‐organic solvent systems. © 2006 Wiley Periodicals, Inc. Int J Chem Kinet 38: 159–165, 2006     

Applications of Hadamard transform-gas chromatography/mass spectrometry to online detection of exhaled breath after drinking or smoking

A Hadamard transform-gas chromatography/mass spectrometry (HT-GC/MS) technique was employed for the online detection of ethanol or toluene in exhaled breath after drinking or smoking, respectively. Exhaled breath samples, collected from volunteers, were directly injected into the GC inlet by a Hadamard-injector without any pretreatment. In the case of breath from a drinker, using a conventional single injection, a small ion peak (corresponding to ∼0.1 ng of ethanol), the intensity of which was approximately equal to or less than the limit of detection. When the HT technique was applied, the signal-to-noise (S/N) ratio was dramatically improved. Furthermore, in the case of breath from a smoker, using conventional injection, a weak ion peak (corresponding to ∼0.7 pg of toluene) was marginally detected. However, the HT technique led to an improvement in the S/N ratio, with the peak corresponding to the limit of detection. In both cases, the HT technique permitted specific components in exhaled breath to be determined, without the need for any extraction procedures.     

Volatile Organic Compounds Analysis in Breath Air in Healthy Volunteers and Patients Suffering Epidermoid Laryngeal Carcinomas

Exhaled breath contains thousands of gaseous volatile organic compounds (VOCs) that may be used as non-invasive markers of head and neck epidermoid cancer. We hypothesized that solid phase micro-extraction coupled to gas chromatography–mass spectrometry can discriminate patients with epidermoid head and neck cancer from healthy controls by analyzing the gaseous volatile organic compounds, VOC-profile, in exhaled breath, thus identifying some non-invasive biomarkers to be used in early detection. Twenty healthy subjects participated in a cross-sectional study plus 11 patients with epidermoid supraglottic laryngeal cancer. VOCs from T3 supraglottic cancer were clustered distinctly from those of T1 and healthy subjects. Up to seven VOCs were detected differently from healthy volunteers, mainly 2-butanone and ethanol. Thus VOC-patterns of exhaled breath may discriminate patients with epidermoid head and neck cancer from healthy controls.     

Monitoring chloramines and bromamines in a humid environment using selected ion flow tube mass spectrometry

The selectivity and sensitivity of selected ion flow tube mass spectrometry (SIFT‐MS) for individual breath analysis of haloamines has been improved by heating the flow tube in a commercial instrument to around 106°C. Data is presented showing the marked reduction in the number density of water clusters of product ions of common breath metabolites that are isobaric with the product ions from monochloramine and monobromamine that are used to monitor the haloamine concentrations. These results have direct relevance to the real‐time monitoring of chloramines in drinking water, swimming pools and food processing plants. However, once the isobaric overlaps from water cluster ions are reduced at the higher temperatures, there is no conclusive evidence showing the presence of haloamines on single breath exhalations in the mid parts per trillion range from examination of the breaths of volunteers. Copyright © 2010 John Wiley & Sons, Ltd.     

Acetone,butanone, pentanone,hexanone and heptanone in the headspace of aqueous solution and urine studied by selected ion flow tube mass spectrometry

Urine is commonly analysed in clinical practice by a variety of liquid‐phase techniques to check for excessive ketone bodies, proteins and salts to name just a few compounds. However, little work has been carried out to measure the volatile compounds emitted by urine since these do not yet have an established role in clinical diagnosis. There is, however, a growing body of evidence that these volatile compounds can be indicators of adverse physiological conditions and disease and with the advent of sensitive gas‐phase analytical methods they can be quickly quantified in urine headspace and potentially provide valuable support for clinical diagnosis. Thus, we are developing selected ion flow tube mass spectrometry, SIFT‐MS, for the real‐time analysis of urine headspace, ultimately to support rapid diagnosis in the clinical environment. In this paper we focus on volatile ketones in the headspace of aqueous solutions and urine donated by three healthy volunteers. Using SIFT‐MS, we have unambiguously quantified in urine headspace acetone, by far the most abundant ketone, butanone, pentanone, hexanone and heptanone using NO⁺ precursor ions. Further to this, we have determined the Henry's Law coefficients, HLC, for these ketones in aqueous solution to allow the liquid‐phase concentrations in urine to be estimated from headspace levels of their vapours. In addition, the influence of the addition of physiological amounts of dissolved urea, sodium chloride and hydrochloric acid on the partitioning of these ketones between the aqueous phase and gas phase has been investigated and found to be small, which gives greater credence to the use of the HLC obtained using aqueous solutions for the estimation of ketone concentrations in urine. Finally, parallel measurements of the levels of acetone in exhaled breath and urine headspace have been obtained and shown to be very similar, which gives support to the previous deduction from breath analysis that acetone is a truly systemic compound. Copyright © 2009 John Wiley & Sons, Ltd.     

Molecular Solution Behaviour of an Intermediate Biofuel Feedstock: Acetone–Butanol–Ethanol (ABE)

Mixtures of acetone, butanol, and ethanol (ABE) are common intermediate products in the production of biofuels via biomass fermentation. Their separation to yield, for example, bio‐butanol, is still difficult due to the lack of a fundamental understanding of these mixtures at the molecular level. In order to bridge this gap, a detailed analysis of characteristic features of the vibrational spectrum is carried out. A systematic study of the binary solutions of acetone with ethanol and butanol does not only reveal a universal behaviour at the molecular level when acetone is mixed with short‐chain alcohols, it also shows that the phenomena at a length scale between the molecules and in the macroscopic solution need to be taken into account to understand the structure–property relationships. The size of self‐associated molecule clusters seems to determine whether or not a system exhibits an azeotrope. When a second alcohol is added to an acetone/alcohol solution, no additional non‐idealities are induced, which is advantageous for modelling ternary ABE mixtures and for improving their processing in the production of biofuels.     

Analysis of exhaled breath from smokers,passive smokers and non‐smokers by solid‐phase microextraction gas chromatography/mass spectrometry

In this study, 38 samples of expired air were collected and analyzed from 20 non‐smoking volunteers, four passive smokers and 14 smokers (21 women and 17 men). Measurements were carried out using solid‐phase microextraction (SPME) as an isolation and preconcentration technique. The determination and identification were accomplished by gas chromatography coupled with mass spectrometry (GC/MS). Our data showed that ca 32% of all identified compounds in the breath of healthy non‐smokers were saturated hydrocarbons. In the breath of smoking and passive smoking volunteers hydrocarbons were predominant, but also present were more exogenous analytes such as furan, acetonitrile and benzene than in the breath of non‐smokers. Acetonitrile, furan, 3‐methylfuran, 2,5‐dimethylfuran, 2‐butanone, octane and decane were identified in breath of smoking and passive smoking persons. Copyright © 2008 John Wiley & Sons, Ltd.     

选择离子流动管质谱及其在痕量气体分析中的应用

选择离子流动管质谱(SIFT-MS)结合流动管技术、化学电离和质谱,有选择地使用F13O^ 、NO^ 和O2^ 初始离子,可在几秒之内对空气、呼吸气体和液表蒸气中的痕量气(如乙醇、乙醛、丙酮、氨和2-甲基丁二烯等,行多组分实时在线分析。介绍了选择离子流动管(SIFT)技术、SIFT-MS的分析方法及其物理和离子化学基础、SIFT-MS在不同领域的痕量气体分析中的应用。