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.     

Repeatability of the measurement of exhaled volatile metabolites using selected ion flow tube mass spectrometry

Selected ion flow tube mass spectrometry, SIFT-MS, has been used to determine the repeatability of the analysis of volatile metabolites within the breath of healthy volunteers, with emphasis on the influence of sampling methodology. Baseline instrument specific coefficients of variability for examined metabolites were as follows: acetone (1%), ammonia (1%), isoprene (2%), propanol (6%), ethanol (7%), acetic acid (7%), and hydrogen cyanide (19%). Metabolite concentration and related product ion count rate were identified as strong determinants of measurement variation. With the exception of ammonia, an orally released metabolite, variability in repeated on-line breath analysis tended to be lower for metabolites of systemic origin. Standardization of sampling technique improved the repeatability of the analysis of selected metabolites. Off-line (bag) alveolar breath sampling, as opposed to mixed (whole) breath sampling, likewise improved the repeatability of the analysis of all metabolites investigated, with the exception of acetic acid. We conclude that SIFT-MS analysis of common volatile metabolites within the breath of healthy volunteers is both reliable and repeatable. For selected metabolites, the finding that repeatability is improved through modification of sampling methodology may have implications in terms of future recommended practices.     

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).     

Quantification of hydrogen cyanide in humid air by selected ion flow tube mass spectrometry

Following our recent observation that Pseudomonas bacteria in vitro emit hydrogen cyanide, we have found it necessary to investigate the ion chemistry of this compound and to extend the kinetics database for selected ion flow tube mass spectrometry (SIFT-MS) to allow the accurate quantification of HCN in moist air samples, including exhaled breath. Because of the proximity of the proton affinities of HCN and H2O molecules, the presence of water vapour can significantly distort HCN analysis in the presence of water vapour and a more sophisticated analytical procedure has to be developed. Thus, the reactions of H3O+(H2O)0,1,2,3 ions with HCN molecules have been studied in the presence of varying concentrations of water vapour, reactions on which SIFT-MS analysis of HCN relies. The results of these experiments have allowed an analytical procedure to be developed which has extended the kinetics database of SIFT-MS, such that HCN can now be quantified in humid air and in exhaled breath.     

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。     

Quantification of volatile compounds in the headspace of aqueous liquids using selected ion flow tube mass spectrometry

We describe a method by which the concentrations of volatile compounds in the headspace of their dilute aqueous solutions in sealed containers can be determined using on-line selected ion flow tube mass spectrometry (SIFT-MS). Thus, the changing number density of the molecules of the volatile compound in the carrier gas of the SIFT-MS instrument is described in terms of its changing flow rate as the pressure in the sealed container decreases during the sampling procedure. It is shown that the best analytical procedure is to determine the mean concentration of the trace gas in the liquid headspace over a given sampling time and relate this to the required concentration, which is the initial equilibrium concentration established before the pressure in the sealed container reduces significantly. To test the validity of this analytical approach, the headspace concentrations of acetaldehyde, ethanol and acetone above aqueous solutions of known concentrations have been determined. Hence, the Henry's Law constants for these compounds have been determined and found to agree with the published values. The confirmation of the quality of this sampling methodology combined with SIFT-MS for the analysis of volatile compounds in liquid headspace paves the way for the rapid analyses of biological liquids such as urine and serum for clinical diagnosis and physiological monitoring.     

Time-resolved selected ion flow tube mass spectrometric quantification of the volatile compounds generated by E. coli JM109 cultured in two different media

Preliminary measurements have been made of the volatile compounds emitted by the bacterium E. coli JM109 cultured in the commonly used media Dulbecco's modified Eagle's medium (DMEM) and lysogeny broth (LB) using selected ion flow tube mass spectrometry, SIFT-MS, as a step towards the real time, non-invasive monitoring of accidental infections of mammalian cell cultures. In one procedure, the culture medium alone and the E. coli cells/medium combination were held at 37 °C in bottles sealed with septa for a given time period, usually overnight, to allow the bacterium to proliferate, after which the captured headspace was analysed directly by SIFT-MS. Several compounds were seen to be produced by the E. coli cells that depended on the liquid medium used: when cultured in DMEM, copious amounts of ethanol, acetaldehyde and hydrogen sulphide were produced; in LB ammonia is the major volatile product. In a second procedure, to ensure aerobic conditions prevailed in the cell culture, selected volatile compounds were monitored by SIFT-MS in real time for several hours above the open-to-air E. coli/DMEM culture held at close to 37 °C. The temporal variations in the concentrations of some compounds, which reflect their production rates in the culture, indicate maxima. Thus, the maxima in the ethanol and acetaldehyde production are a reflection of the reduction of glucose from the DMEM by the vigorous E. coli cells and the maximum in the hydrogen sulphide level is an indication of the loss of the sulphur-bearing amino acids from the DMEM. Serendipitously, emissions from DMEM inadvertently infected with the bacterium C. testosteroni were observed when large quantities of ammonia were seen to be produced. The results of this preliminary study suggest that monitoring volatile compounds might assist in the early detection of bacterial infection in large-scale bioreactors.     

Method for the collection and analysis of volatile compounds in the breath

A new method is described for the collection and assay of volatile compounds in the breath. Subjects expired into a pump-assisted collecting apparatus in which the breath was drawn through a water trap and then through an adsorptive trap where the volatile compounds were captured on graphitized carbon and molecular sieve. The sample was subsequently eluted from the trap by thermal desorption, concentrated by two-stage cryofocusing, then assayed by gas chromatography with flame ionization and flame photometric detection. Several compounds were regularly observed in the breath of normal human volunteers, including peaks eluting with the same retention times as isoprene, ethanol, acetone, acetaldehyde and carbon disulfide. As a quantitative assay for endogenous isoprene in the breath, the method was sensitive, linear, accurate and reproducible. This method provided a number of advantages: the collection technique was acceptable to volunteers and could be used at sites remote from the laboratory. The automated assay allowed isoprene and several other volatile compounds in the breath to be observed consistently and with improved sensitivity.     

Analysis of petrol and diesel vapour and vehicle engine exhaust gases using selected ion flow tube mass spectrometry

We have used selected ion flow tube mass spectrometry (SIFT-MS) to analyse the vapours emitted by petrol and diesel fuels and the exhaust gases from petrol (spark ignition) and diesel (compression ignition) engine vehicles fitted with catalytic converters. Only those components of these media that have significant vapour pressures at ambient temperatures were analysed and thus particulates were obviously not detected. These media have been analysed using the full scope of SIFT-MS, i.e., with the three available precursor ions H3O+, NO+ and O2+. The combination of the H3O+ and NO+ analyses is seen to be essential to distinguish between different product ions at the same mass-to-charge ratio (m/z) especially in identifying aldehydes in the exhaust gases. The O2+ precursor ions are used to detect and quantify the large amount of nitric oxide present in the exhaust gases from both engine types. The petrol and diesel vapours consist almost exclusively of aliphatic alkanes, alkenes and alkynes (and dienes) and aromatic hydrocarbons. Some of these compounds appear in the exhaust gases together with several aldehydes, viz. formaldehyde, acetaldehyde, pentanal, pentenal (acrolein), butenal, and also methanol and ethanol. Acetone, nitric oxide and ammonia are also present, acetone and nitric oxide being much more abundant in the diesel exhaust gas than in the petrol exhaust gas. These data were obtained from samples collected into pre-evacuated stainless steel vessels. Trapping of the volatile compounds from the gas samples is not required and analysis was completed a few minutes later. All the above compounds are detected simultaneously, which demonstrates the value of SIFT-MS in this area of research.     

On-line, real time monitoring of exhaled trace gases by SIFT-MS in the perioperative setting: a feasibility study

A study is described of the first on line, real time analyses of the exhaled breath of five anaesthetized patients during the complete perioperative periods of laparoscopic surgery. These breath analyses were achieved using a selected ion flow tube, SIFT-MS, instrument, located in the operating theatre at an acceptable distance from the operating table, and coupled to the endotracheal tube in the ventilation circuit via a 5 metre long capillary tube. Thus, inhalation/exhalation breathing cycles, set to be at a frequency of 10 per minute, were sampled continuously for water vapour, the metabolites acetone and isoprene and the propofol used to induce anaesthesia for each operating period that ranged from 20 min (shortest) to 80 min (longest). Whilst there was some loss of water vapour along the long sampling line, the concentrations of the other trace compounds were not diminished. The breath acetone was essentially at a constant level for each patient, but increased somewhat over the longest operating period due to the onset of lipolysis. Most interesting is the clear increase of breath isoprene following abdomen inflation with carbon dioxide. The vapour of the intravenously injected propofol was detected in the exhaled breath and remained essentially constant during the perioperative period. These analyses were performed totally non-invasively and the data were immediately and constantly available to the anaesthetist and surgeon. Exploitation of this development could influence decision making and potentially improve patient safety within the perioperative setting.     

Influence of water vapour on selected ion flow tube mass spectrometric analyses of trace gases in humid air and breath

Selected ion flow tube mass spectrometry (SIFT-MS) detects and quantifies in real time the trace gases, M, in air/breath samples introduced directly into a flow tube. Inevitably, relatively large partial pressures of water vapour are introduced with the sample and the water molecules become involved in the ion chemistry on which this analytical technique depends. When H(3)O(+) ions are used as the precursors for chemical ionisation and SIFT mass spectrometric analyses of M, they generally result in the formation of MH(+) ions. Also, when water vapour is present the H(3)O(+) ions are partially converted to hydrated hydronium ions, H(3)O(+).(H(2)O)(1,2,3). The latter may act as precursor ions and produce new product ions like MH(+).(H(2)O)(1,2,3) via ligand switching and association reactions. This ion chemistry and the product ions that result from it must be accounted for in accurate analyses by SIFT-MS. In this paper we describe the results of a detailed SIFT study of the reactions involved in the quantification of acetone, ethyl acetate, diethyl ether, methanol, ethanol, ammonia and methyl cyanide by SIFT-MS in the presence of water vapour. This study was undertaken to provide the essential data that allows more accurate analyses of moist air and breath by SIFT-MS to be achieved. It is shown using our standard analysis procedure that the error of SIFT-MS quantification caused by the presence of water vapour is typically 15%. An improved analysis procedure is then presented that is shown to reduce this error to typically 2%. Additionally, some fundamental data have been obtained on the association reactions of protonated organic molecules, MH(+) ions, with water molecules forming MH(+).H(2)O monohydrate ions. For some types of M, reaction sequences occur that lead to the formation of dihydrate and trihydrate ions.     

异戊二烯与OH自由基光化学反应的二次有机气溶胶的生成

利用烟雾箱模拟装置,研究了异戊二烯与OH自由基反应的二次有机气溶胶(SOA)的生成.反应中生成的气相产物通过质子转移反应质谱仪(PTR-MS)测定,SOA的浓度及粒径谱分布通过高分辨率粒径谱仪(EEPS3090)测定.研究表明:甲基丙烯醛(MAC)/甲基乙烯基酮(MVK)、乙醛、甲醛、甲醇、甲酸/乙醇、乙醇醛、甲基乙二醛、丙酮/丙醛等为主要气相产物,各组实验中MAC/MVK和乙醛浓度达到最大时,其产率分别介于13.78%～37.72%和5.38%～9.34%(以C计)范围内;SOA生成量及其中值粒径随异戊二烯反应量的增加而增加,气相物质稳定后,SOA产率在5.6%～11.7%范围内,粒径在22～165nm范围内.     

Kinetics of ethanol decay in mouth‐ and nose‐exhaled breath measured on‐line by selected ion flow tube mass spectrometry following varying doses of alcohol

A study has been carried out of the decay of ethanol in mouth‐exhaled and nose‐exhaled breath of two healthy volunteers following the ingestion of various doses of alcohol at different dilutions in water. Concurrent analyses of sequential single breath exhalations from the two volunteers were carried out using selected ion flow tube mass spectrometry, SIFT‐MS, on‐line and in real time continuously over some 200 min following each alcohol dose by simply switching sampling between the two volunteers. Thus, the time interval between breath exhalations was only a few minutes, and this results in well‐defined decay curves. Inspection of the mouth‐exhaled and nose‐exhaled breath data shows that mouth contamination of ethanol diminished to insignificant levels after a few minutes. The detailed results of the analyses of nose‐exhaled breath show that the peak levels and the decay rates of breath ethanol are dependent on the ethanol dose and the volume of ethanol/water mixture ingested. From these data, both the efficiency of the first‐pass metabolism of ethanol and the indications of gastric emptying rates at the various doses and ingested volumes have been obtained for the two volunteers. Additionally and simultaneously, acetaldehyde, acetic acid and acetone were measured in each single breath exhalation. Acetaldehyde, the primary product of ethanol metabolism, is seen to track the breath ethanol. Acetic acid, a possible secondary product of this metabolism, was detected in the exhaled breath, but was shown to largely originate in the oral cavity. Breath acetone was seen to increase over the long period of measurement due to the depletion of nutrients. Copyright © 2010 John Wiley & Sons, Ltd.     

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.     

Selected ion flow tube mass spectrometry for on-line trace gas analysis in biology and medicine

Selected ion flow tube mass spectrometry, (SIFT-MS), is a technique for simultaneous real-time quantification of several trace gases in air and exhaled breath. It relies on chemical ionization of the trace gas molecules in air/breath samples introduced into helium carrier gas, using H(3)O(+), NO(+) and O(2)(+) reagent (precursor ions). Reactions between the precursor ions and the trace gas molecules proceed for an accurately defined time, the precursor and product ions being detected and counted by a downstream mass spectrometer. Absolute concentrations of trace gases in single breath exhalation can be determined by SIFT-MS down to parts-per-billion (ppb) levels, obviating sample collection into bags or onto traps. Calibration using chemical standards is not required, as the concentrations are calculated using the known reaction rate constants and measured flow rates and pressures. SIFT-MS has been used for many pilot investigations in several areas of research, especially as a non-invasive breath analysis tool to investigate physiological processes in humans and animals, for clinical diagnosis and for therapeutic monitoring. Examples of the results obtained from several such studies are outlined to demonstrate the potential of SIFT-MS for trace gas analysis of air, exhaled breath and the headspace above liquids.     

Development and validation of a direct headspace GC-FID method for the determination of sevoflurane, desflurane and other volatile compounds of forensic interest in biological fluids: application on clinical and post-mortem samples

A simple and reliable headspace GC‐flame ionization detection (HS‐GC‐FID) method has been developed and validated for the simultaneous determination of seven volatile compounds of forensic interest: sevoflurane, desflurane, ethanol, methanol, 1‐propanol, acetone and acetaldehyde. All seven compounds including acetonitrile (internal standard) eluted within 10 min and were well resolved with no endogenous interference. Good linearity was observed in the range of 1–12 mg/dL for both anesthetics and 2.5–40 mg/dL for the other five analytes. The method showed good precision, sensitivity and repeatability. Most of the analytes remained stable during the storage of samples at 4°C. Desflurane and acetone degraded (>10%), when the samples remained on the autosampler for more than 2 and 3 h, respectively. The method was finally applied on clinical and post‐mortem blood and urine samples. The clinical samples were collected both from patients who underwent surgery, as well as from the occupationally exposed medical and nursing staff of the university hospital, working in the operating rooms. The hospital staff samples were found negative for all compounds, while the patients' samples were found positive for the anesthetic administered to the patient. The post‐mortem blood samples were found positive for ethanol and acetaldehyde.     

Combined use of gas chromatography and selected ion flow tube mass spectrometry for absolute trace gas quantification

The value of the gas chromatography (GC) and selected ion flow tube mass spectrometry (SIFT-MS) combination for the analysis of trace gases is demonstrated by the quantification of acetone in air samples using the three precursor ions available to SIFT-MS, viz. H3O+, NO+ and O2+, and by the separation of the isomers 1-propanol and 2-propanol, and their analysis using H3O+ precursor ions. It is shown that the GC/SIFT-MS combination allows for accurate trace gas quantification obviating the regular, time-consuming calibrations that are usually required for the more commonly used detectors of GC systems, and the positive identification of isomers in mixtures that is often challenging using SIFT-MS alone. Thus, the GC/SIFT-MS combination paves the way to more confident analyses of complex mixtures such as exhaled breath.     

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.     

Electrical sensing of volatile organic compounds in exhaled breath for disease diagnosis

In recent years, electrical sensors toward breath volatolomics have attracted increasing interest owing to their wide feasibility in noninvasive disease diagnostics. In this article, the working principles of active nanomaterials (e.g. metal oxides, polymers, and nanocarbon) toward volatile organic compounds are presented, with a special focus on the influence of surface chemistry and structural feature of these nanomaterials on the sensing performance. The latest and representative achievements on the direct analysis of three typical exhaled volatile organic compounds, including acetone, ammonia, and hydrogen sulfide, that are recognized as important disease biomarkers, are highlighted, indicating the capability of the electrical sensors in enabling noninvasive diagnosis and real-time monitoring. The opportunities and challenges in this field are provided in the end, with an emphasis on the background interference and data recognition which are key factors in developing prospective electrical sensors toward volatolomics analysis.     

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.