Non-invasive disease monitoring on the basis of volatile breath markers is a very attractive but challenging task. Several hundreds of compounds have been detected in exhaled air using modern analytical techniques (e.g. proton-transfer reaction mass spectrometry, gas chromatography-mass spectrometry) and have even been linked to various diseases. However, the biochemical background for most of compounds detected in breath samples has not been elucidated; therefore, the obtained results should be interpreted with care to avoid false correlations. The major aim of this study was to assess the effects of smoking on the composition of exhaled breath. Additionally, the potential origin of breath volatile organic compounds (VOCs) is discussed focusing on diet, environmental exposure and biological pathways based on other's studies. Profiles of VOCs detected in exhaled breath and inspired air samples of 115 subjects with addition of urine headspace derived from 50 volunteers are presented. Samples were analyzed with GC-MS after preconcentration on multibed sorption tubes in case of breath samples and solid phase microextraction (SPME) in the case of urine samples. Altogether 266 compounds were found in exhaled breath of at least 10% of the volunteers. From these, 162 compounds were identified by spectral library match and retention time (based on reference standards). It is shown that the composition of exhaled breath is considerably influenced by exposure to pollution and indoor-air contaminants and particularly by smoking. More than 80 organic compounds were found to be significantly related to smoking, the largest group comprising unsaturated hydrocarbons (29 dienes, 27 alkenes and 3 alkynes). On the basis of the presented results, we suggest that for the future understanding of breath data it will be necessary to carefully investigate the potential biological origin of volatiles, e.g., by means of analysis of tissues, isolated cell lines or other body fluids. In particular, VOCs linked to smoking habit or being the results of human exposure should be considered with care for clinical diagnosis since small changes in their concentration profiles (typically in the * This work was presented at

Isoprene concentrations in exhaled breath showed gender-specific correlations with respect to age. Further investigations are necessary to clarify the relation between isoprene concentrations in exhaled breath and cholesterol levels and synthesis rates in blood.

The present study was performed to determine the variations of breath acetone concentrations with age, gender and body-mass index (BMI). Previous investigations were based on a relatively small cohort of subjects (see Turner et al 2006 Physiol. Meas. 27 321-37). Since exhaled breath analysis is affected by considerable variation, larger studies are needed to get reliable information about the correlation of concentrations of volatiles in breath when compared with age, gender and BMI. Mixed expiratory exhaled breath was sampled using Tedlar bags. The concentrations of a mass-to-charge ratio (m/z) of 59, attributed to acetone, were then determined using proton transfer reaction-mass spectrometry. Our cohort, consisting of 243 adult volunteers not suffering from diabetes, was divided into two groups: one that fasted overnight prior to sampling (215 volunteers) and the other without a dietary control (28 volunteers). In addition, we considered a group of 44 healthy children (5-11 years old).The fasted subjects' concentrations of acetone ranged from 177 ppb to 2441 ppb, with an overall geometric mean (GM) of 628 ppb; in the group without a dietary control, the subjects' concentrations ranged from 281 ppb to 1246 ppb with an overall GM of 544 ppb. We found no statistically significant shift between the distributions of acetone levels in the breath of males and females in the fasted group (the Wilcoxon-Mann-Whitney test yielded p = 0.0923, the medians being 652 ppb and 587 ppb). Similarly, there did not seem to be a difference between the acetone levels of males and females in the group without a dietary control. Aging was associated with a slight increase of acetone in the fasted females; in males the increase was not statistically significant. Compared with the adults (a merged group), our group of children (5-11 years old) showed lower concentrations of acetone (p < 0.001), with a median of 263 ppb. No correlation was found between the acetone levels and BMI in adults. Our results extend those of Turner et al's (2006 Physiol. Meas. 27 321-37), who analyzed the breath of 30 volunteers (without a dietary control) by selected ion flow tube-mass spectrometry. They reported a positive correlation with age (but without statistical significance in their cohort, with p = 0.82 for males and p = 0.45 for females), and, unlike us, arrived at a p-value of 0.02 for the separation of males and females with respect to acetone concentrations. Our median acetone concentration for children (5-11 years) coincides with the median acetone concentration of young adults (17-19 years) reported by Spanel et al (2007 J. Breath Res. 1 026001).

. tumour necrosis factoralpha plasma level in patients with type 1 diabetes mellitus and its association with glycaemic control and cardiovascular risk factors. J Intern Med 2000: 248: 67±76.Objectives. Diabetic patients reveal a significant increase in their cardiovascular risk. Beside glycaemic control and management of established risk factors, determination of cytokines, like serum levels of tumour necrosis factor-alpha (TNF-a), might offer a tool to determine patients at high risk. The cytokine TNF-a reveals a complex relationship with diabetes. It is involved in beta-cell damage leading to type 1 diabetes, causes insulin resistance associated with obesity and is of influence in the formation of atherosclerotic vascular lesions. We were interested in the possible association of this cytokine with metabolic control and cardiovascular risk factors in patients with type 1 diabetes. Design and Subjects. TNF-a plasma levels were determined in 44 outdoor patients (15 women, 29 men) with type 1 diabetes mellitus (mean duration 11.2 6 8.7 years) and in 24 healthy controls by use of a solid phase enzyme amplified sensitivity immunoassay (TNF-a ELISA, Biosource Fleurus, Belgium). None of our study participants suffered from inflammatory or other concurrent diseases. Relationships between variables were evaluated by non-parametric Spearman correlation coefficients.Results. TNF-a plasma levels were significantly higher in diabetic patients (19.3 6 7.5 pg mL 21 ) than in non-diabetic subjects (11.1 6 5.8 pg mL 21 ; P , 0.023), and revealed a significant positive correlation with glycated haemoglobin (HbA 1c ) (r = 0.43; P , 0.004) and fructosamine (r = 0.31; P , 0.049) values, and a negative correlation with HDL cholesterol (r = ±0.36; P , 0.018) and apoAIlevels (r = ±0.37; P , 0.015). These relationships could be observed in patients with a duration of diabetes for more than 5 years, as well as in patients with a shorter duration of diabetes. In the male group, TNF-a plasma levels revealed a significant positive correlation with plasma levels of thiobarbituric acid reacting substances (r = 0.61; P , 0.001). Plasma levels of thiobarbituric acid reacting substances showed a positive correlation with the duration of diabetes (r = 0.58; P , 0.008), as well as with the serum levels of the vascular adhesion molecules intercellular adhesion molecule (ICAM) (r = 0.34; P , 0.051) and vascular cell adhesion molecule (VCAM) (r = 0.30; P , 0.052). Conclusions. Our data indicate that TNF-a plasma levels are increased in type 1 diabetes mellitus and reveal a significant association with metabolic longterm control parameters, HbA 1c and fructosamine for glycaemic control, and HDL cholesterol for triglyceride metabolism, as well with lipid peroxidation.

A pilot study has been carried out to define typical characteristics of the trace gas compounds in exhaled breath of non-smokers and smokers to assist interpretation of breath analysis data from patients who smoke with respiratory diseases and lung cancer. Exhaled breath was analyzed using proton transfer reaction-mass spectrometry (PTR-MS) for 370 volunteers (81 smokers, 210 non-smokers, 79 ex-smokers). Volatile organic compounds corresponding to product ions at seven mass-to-charge ratios (m/z 28, 42, 69, 79, 93, 97, 123) in the PTR-MS spectra differentiated between smokers and non-smokers. The Youden index (= maximum of sensitivity + specificity - 1, YI) as a measure for differentiation between smokers and non-smokers was YI = 0.43 for ions at the m/z values 28 (tentatively identified as HCN), YI = 0.75 for m/z = 42 (tentatively identified as acetonitrile) and YI = 0.53 for m/z = 79 (tentatively identified as benzene). No statistically significant difference between smokers and non-smokers was observed for the product ions at m/z = 31 and 33 (compounds tentatively identified as formaldehyde and methanol). When interpreting the exhaled breath of lung cancer or COPD patients, who often smoke, compounds appearing at the above-mentioned seven mass-to-charge ratios should be considered with appropriate care to avoid misdiagnosis. Validation studies in larger numbers of patients with more precise delineation of their smoking behavior and using additional analytical techniques such as GC/MS and SIFT-MS should be carried out.