Short Communication Received: 17 June 2008, Revised: 14 August 2008, Accepted: 15 August 2008 Published online 27 November 2008 in Wiley Interscience (www.interscience.wiley.com) DOI 10.1002/bmc.1141 Biomed. Chromatogr. 2009; 23: 551– 556 Copyright © 2008 John Wiley & Sons, Ltd. 551 John Wiley & Sons, Ltd. Analysis of exhaled breath from smokers, passive smokers and non-smokers by solid- phase microextraction gas chromatography/ mass spectrometry Analysis of exhaled breath by SPME GC/MS BogusÝaw Buszewski, a * Agnieszka Ulanowska, a,b Tomasz Ligor, a,b Natalia Denderz a and Anton Amann b ABSTRACT: 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. Keywords: breath analysis; lung diseases; cigarette smoking; biomarkers Introduction The role of tobacco smoking and involuntary smoking in cancer is very well documented in the scientific literature and was recently re-evaluated by the International Agency for Research on Cancer (IARC). The recent report concludes that tobacco smoking and tobacco smoke are carcinogenic to humans (IARC Group 1 classi- fication) and that involuntary smoking (exposure to second-hand or ‘environmental’ tobacco smoke) is carcinogenic to humans (Group 1; Lewtas, 2007). Many nitrogen-containing substances have been found in the mutagenic fraction of cigarette smoke condensate. It is also known that smoking cigarettes increases the amount of free radicals in human body (Gelvan et al., 1995). The excess of free oxygen species causes the disturbance in intracellular balance which is commonly called oxidative stress. In pathological states DNA, other proteins and lipids are dam- aged. These changes in cells and genetic material accelerate the cells’ senescence and have the direct influence in forming many different diseases and also cancers. During metabolic processes occurring in live organisms many substances are generated. These compounds penetrate the bloodstream and circulate through the whole body. The barrier between blood and alveolar air is very thin, which is why metab- olites can be identified in breath samples. Therefore, breath air is a complex mixture of volatile organic compounds (VOCs) (i.e. alcohols, aldehydes, hydrocabons, etc.), non-volatile organic compounds (i.e. leukotrienes and prostaglandins) and inorganic compound such as water vapor, carbon dioxide and nitrogen (Aghdassi and Allard, 2004; Bucchioni et al., 2004; Buszewski et al., 2007). Some of them are recognized as biomarkers. Biomarkers present in breath can be exogenous or endogenous. Benzene, ethylbenzene, xylene isomers, toluene and trichloromethane are supposed to be exogenous and they are probably inhaled with ambient air or ingested with food (Plebani et al., 1999). Endogenous biomarkers are produced in the body during lipid oxidation (e.g. ethane, pentane) or can be a by-product of synthesis for more complicated biomolecules such as cholesterol (Miekisch et al., 2004; Buszewski et al., 2007). Their presence and concentration provide valuable information about human health and can be an indicator of pathological states. For example, pentane, acetal- dehyde, 2-propanol and heptanal were recognized as biomark- ers of breast cancer (Phillips et al., 2006). Other endogenous compounds such as isoprene and acetone and also exogenous * Correspondence to: B. Buszewski, Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin Str., PL-87-100 Toruñ, Poland. E-mail: bbusz@chem.uni.torun.pl a Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin Str., PL-87-100 Toruñ, Poland b Department of Anesthesiology and Critical Medicine Care, Innsbruck Medi- cal University, A-6020 Innsbruck, Austria Abbreviations used: COPD, chronic obstructive pulmonary disease; IARC, International Agency for Research on Cancer; VOCs, volatile organic compounds. Contract/grant sponsor: European Commission; Contract/grant number: BAMOD 19031. Contract/grant sponsor: Polish Ministry of Sciences and High Education; Contract/grant number: N204 165 31/3730. Contract/grant sponsor: CEEPUS-II scholarship; Contract/grant number: CII- PL-0004-01-0506-M-4479 and CII-PL-0004-02-0607-M-12188.