Chemotherapy control by breath profile with application of SPME-GC/MS method.

Chemotherapy used as a treatment against lung cancer has influence on metabolic processes occurring in healthy cells. The changes of biochemical pathways proceeded inside cells might be observed in expired air. In the experiment, breath analysis was carried out before and after anticancer therapy. Expired air samples were collected from 22 patients with a biopsy confirmed lung cancer. Volatile organic compounds present in breath were analyzed by gas chromatography/mass spectrometry. For enrichment of analytes solid-phase microextraction technique was applied. Eight fibers covered by different sorbents were tested. Carboxen-polydimethylsiloxane fiber revealed the highest extraction efficiency in relation to analytes in breath. The data showed that cytostatic drugs increase the concentration of acetone and isoprene in the breath collected after chemotherapy. Volatile metabolites of administrated drugs were not identified in expired air.

Evaluation of septa quality for automatic SPME-GC-MS trace analysis.

The vials used for the preparation of breath samples for automated solid-phase microextraction-gas chromatography-mass spectrometry analysis are crimped with septa. These septa often emit specific volatile organic compounds (VOCs) confounding the measurement results of breath samples. In the current paper, 14 different brands of magnetic caps with silicone-polytetrafluoroethylene (PTFE), butyl-PTFE, or butyl rubber septa were tested. The total emission of septa over a 4 h period was also evaluated. The tested septa emitted 39 different compounds, which are mainly hydrocarbons, alcohols, and ketones. Acetone and toluene are the most abundant out-gassing products. The concentration of acetone was in the range from 55 to 694 ppb for butyl-PTFE septum (brand 14) and butyl rubber (brand 10), respectively. The measured toluene amount was 69-1323 ppb for the septum brand 14 and brand 8 (silicone-PTFE), respectively. Generally, the butyl rubber septa released higher amounts of contaminants in comparison to the silicone ones.

Investigation of lung cancer biomarkers by hyphenated separation techniques and chemometrics.

BACKGROUND: Volatile organic compounds (VOCs) present in breath of lung cancer patients and released by lung cancer cells were determined. As a reference sample breath collected from healthy volunteers and lung segments of cells with proper morphology were used. METHODS: Solid phase microextraction and gas chromato-graphy mass spectrometry (SPME-GC/MS) method was applied for VOC analysis. Statistical methods have been developed to find similarities between patterns of investigated compounds within three populations of samples: exhaled air samples from patients with lung cancer, headspace of lung tissues with healthy and cancer cells. RESULTS: Quantitative analysis of VOCs in the headspace of lung tissues revealed that cancer cells released higher concentrations of ethanol, acetone, carbon disulfide, dimethyl sulfide, 1-propanol, 2-propanol, 2-butanone and 2-pentanone than healthy tissues. The increase of concentration of the same compounds was observed in the breath of patients with lung cancer in comparison to breath from healthy non-smoking volunteers. CONCLUSIONS: Detection of lung cancer is possible by volatile biomarkers analysis in breath.

The application of statistical methods using VOCs to identify patients with lung cancer.

In this work, an attempt was made to determine a group of lung cancer biomarkers. For this study, breath samples collected from 137 patients with confirmed lung cancer were analyzed by the SPME-GC/MS method. As a reference group, exhaled air from 143 healthy volunteers with different smoking habits (active smokers, passive smokers and nonsmokers) was applied. Statistical methods such as discriminant analysis (DA) and the CHAID model tree were used for data processing and evaluation. In the breath of patients with lung cancer, increased concentration of ethanol, acetone, butane, dimethyl sulfide, isoprene, propanal, 1-propanol, 2-pentanone, furan, o-xylene and ethylbenzene was observed in comparison to healthy nonsmokers. Furthermore, pentanal, hexanal and nonane were identified only in the breath of people who suffered from cancer. DA confirmed the importance of these compounds and allowed us to identify patients with lung cancer from healthy volunteers. In the exhaled air of healthy smokers (passive and active), a higher concentration of acetonitrile, benzene and furan derivatives was observed than in nonsmokers. DA revealed that in order to recognize healthy volunteers with different smoking habits by breath analysis, butyrolactone, carbon disulfide and dimethyl sulfide have to be considered.

Determination of volatile organic compounds in human breath for Helicobacter pylori detection by SPME-GC/MS.

Helicobacter pylori living in the human stomach release volatile organic compounds (VOCs) that can be detected in expired air. The aim of the study was the application of breath analysis for bacteria detection. It was accomplished by determination of VOCs characteristic for patients with H. pylori and the analysis of gases released by bacteria in suspension. Solid-phase microextraction was applied as a selective technique for preconcentration and isolation of analytes. Gas chromatography coupled with mass spectrometry was used for the separation and identification of volatile analytes in breath samples and bacterial headspace. For data calculation and processing, discriminant and factor analyses were used. Endogenous substances such as isobutane, 2-butanone and ethyl acetate were detected in the breath of persons with H. pylori in the stomach and in the gaseous mixture released by the bacteria strain but they were not identified in the breath of healthy volunteers. The canonical analysis of discrimination functions showed a strong difference between the three examined groups. Knowledge of substances emitted by H. pylori with the application of an optimized breath analysis method might become a very useful tool for noninvasive detection of this bacterium.

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.

Identification of volatile organic compounds secreted from cancer tissues and bacterial cultures.

The early cancer diagnosis increases the possibility of total recovery. The infection of Helicobacter pylori is associated with gastric cancer, the second most common cancer in the world. The determination of volatile organic compounds (VOCs) excreted by stomach tissue and bacteria culture has been investigated. Solid-phase microextraction (SPME) was used for preconcentration and the determination was accomplished by gas chromatography coupled with mass spectrometry (GC/MS). The samples of tissue were taken from five patients (ten samples) with stomach cancer and normal (non-cancerous) segments from other parts of the stomach were used as a control. Eighteen compounds were identified in stomach tissue and seven of them were present both in healthy and cancer tissue. These compounds assumed to be endogenous and acetone ratio (AR) was calculated for ethanol, butane, carbon disulfide, 1-propanol, 2-butanone and 2-pentanone. The data shows that amount of 1-propanol and carbon disulfide in the gaseous composition is higher in cancer tissue than in normal tissue. Eight compounds were identified both in bacteria and tissue. These data suggest that bacteria present in the stomach might cause the increase in the concentration of 1-propanol and carbon disulfide in emission from cancer tissue.