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Acupuncture and Related Therapies 2 (2014) 25–28

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26 J. Friberg / Acupuncture and Related Therapies 2 (2014) 25–28

Volatile organic compounds analyzed by gas chromatography-deep

ultraviolet spectroscopy
Jonas Friberg
JFKemiska, Hyllievångsvägen 17, SE-216 25 Malmö, Sweden

Fig. 1. Schematic illustration of the GC-UV instrumentation.

a r t i c l e i n f o a b s t r a c t
2.2. Sample preparation technique extracts both volatile and non-volatile analytes from different kinds
Article history: of media
Exhaled breath contains thousands such asorganic
of volatile the exhaled breath
compounds collected
(VOCs) in athe
of which Tedlar bag. The
composition varies
In order11
Received toDecember
collect the compounds present in depending
2013 exhaled air, onseveral
status of thequantity
individualof and
the extracted
Differentby the fibreprocesses
metabolic is proportional
within the
technologies are available.
Accepted 11 December 2013 Solid-phase microextraction
body produce(SPME), is substances
volatile to its that
concentration in the
are released into theblood.
sample Whenasthelong
the lungsisthe
a sample preparation technique used both in products the laboratory and into lung
are released reached.
tissue and airways.
on-site. SPME involves the use of a fibre coated with Also,an extracting
chronic inflammation and/or oxidative
After extraction, thestress
SPME can result
fibre in the excretion
is transferred to theofinjection
volatile compounds
port of
Exhaled breath
phase, that can be a liquid (polymer) or a solid (sorbent),
that generate which
unique VOC thepatterns.
gas Therefore, measuring
chromatograph the where
(GC), presencedesorption
of VOCs in exhaled
of the air (breath-
Volatile organic compounds (VOCs)
omics), for clinical diagnosis and monitoring purposes has gained increased interest over the last years.
Deep ultraviolet
This paper describes oneplace
and thebased on gas chromatography
separation is carried out. The(GC) attraction
and deep ultraviolet
Acetone spectroscopy. Spectra of compounds found in exhaled breath are presented.
Benzene © 2013 Elsevier GmbH. All rights reserved.

1. Introduction the volatome, is expected to generate more adequate information

regarding the processes involved. Certainly, analysing the volatome
In ancient times, physicians were aware of the relationship implies a more specific discrimination between various conditions
between the odour of a subjects’ breath and possible diseases as it reflects changes in both exogenous and endogenous com-
associated with it. They realized it could provide insight into phys- pounds [4–9].
iological and pathophysiological processes in the body [1,2]. One technique that allows for the detection of the volatome
It is common knowledge that the sweet acetonic smell of is gas chromatography-deep ultraviolet spectroscopy (GC-DUV). It
breath has been used both for the assessment of environmental factors
might indicate uncontrolled diabetes whereas a fishy reek of breath as well as for individual VOCs [10–17]. However, its use has been
relates to liver disease and a urine-like smell is associated with kid- hampered by its technical design. In the present study I report on
ney failure [3]. Apparently, there is something present in breath a new design that might allow for the use of GC-DUV on a larger
that might enable diagnosing certain diseases. Due to the great scale.
potential of applications in clinical diagnostics and its non-invasive
nature, exhaled air analysis has become of increased interest in
recent years. 2. Materials and methods
Exhaled air breath analysis (breathomics) can be applied as an
analytical and monitoring tool. In the analytical perspective, VOCs 2.1. Sampling of breath
may be used as biomarkers of oxidative stress, inflammation or
carcinogenesis [1]. As monitoring tool, breathomics can be applied Exhaled air comprises a mixture of dead-space air and alveolar
to elucidate the heterogeneity observed in diseases, to study the air. The dead-space air consists of roughly 150 mL air from the upper
pathogen responsible for an infection and to monitor treatment airway where no gaseous exchange between blood and breath air is
efficacy. facilitated [1,4]. Consequently, this part of the exhaled air displays a
The use of individual VOCs as biomarkers of exposure or high resemblance with the previously inspired air. In contrast, alve-
disease is hampered by the fact that using a single compound is olar air originates from the lower airways where gaseous
gener-ally insufficient to monitor complex and heterogeneous exchange between blood and breath air results in concentrations
processes including environmental exposures or chronic of endoge-nous compounds that are two to three times higher
diseases. There-fore, exploring the presence and relations of compared to those observed in dead-space air. Measuring VOCs
exhaled VOCs, called in mixed air implies sampling whole breath that consists of
both dead-space air and alveolar air. An advantage of sampling
E-mail address: 5 L instead of one
single breath is a higher reproducibility and lower variability.
2211-7660/$ – see front matter © 2013 Elsevier GmbH. All rights reserved.
J. Friberg / Acupuncture and Related Therapies 2 (2014) 25–28 27

Fig. 2. (a and b) Schematic presentation of the principles of UV spectroscopy.

28 J. Friberg / Acupuncture and Related Therapies 2 (2014) 25–28

isomers. Selective analysis can be carried out for most of chemical [2] Ma W, Liu X, Pawliszyn J. Analysis of human breath with micro extraction
classes, such as aliphatic aldehydes and ketones, pyridines, ben- techniques and continuous monitoring of carbon dioxide concentration. Anal
Bioanal Chem 2006;385:1398–408.
zene, naphthalenes and higher aromatics, etc. In addition to organic [3] Libardoni M, Stevens PT, Waite JH, Sacks R. Analysis of human breath sam-
compounds also inorganic can be determined with high sensitivity ples with a multi-bed sorption trap and comprehensive two-dimensional gas
[10–17]. UV-spectra in the vapour phase are well defined, which is chromatography (GC × GC). J Chromatogr B 2006;842:13–21.
[4] Miekisch W, Schubert JK, Noeldge-Schomburg GF. Diagnostic potential of
the bases for the identification. breath analysis – focus on volatile organic compounds. Clin Chim Acta
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tion on the collection of exhaled breath in humans. J Appl Physiol 2004;(96):
Data generated by GC-DUV have to be pre-processed properly [6] Schubert JK, Miekisch W, Geiger K, Noldge-Schomburg GF. Breath analy-
in order to obtain more comparable data that can subsequently sis in critically ill patients: potential and limitations. Expert Rev Mol Diagn
be used for data analysis. Similar to other analytical platforms, [7] Buszewski B, Kesy M, Ligor T, Amann A. Human exhaled air analytics: biomark-
breathomic data typically involve a number of pre-processing steps ers of diseases. Biomed Chromatogr 2007;21:553–66.
including noise reduction, background (baseline) correction, align- [8] Phillips M. Method for the collection and assay of volatile organic compounds
in breath. Anal Biochem 1997;247:272–8.
ment and normalization [1,2], Fig. 3a and b.
[9] Van Berkel JJ, Dallinga JW, Moller GM, Godschalk RW, Moonen E, Wouters EF,
Breathomic data display nonlinear parameter dependences. Van Schooten FJ. Development of accurate classification method based on the
Therefore many nonlinear techniques that have been developed analysis of volatile organic compounds from human exhaled air. J Chromatogr
B 2008;2008(861):101–7.
including random forests and two kernel-based models [1,2].
[10] Hatzinikolaou DG, Lagesson V, Stavridou AJ, Pouli AE, Lagesson-Andrasko
L, Stavrides JC. Analysis of the gas phase of cigarette smoke by gas chro-
3. Discussion matography coupled with UV-diode array detection. Anal Chem 2006;78(13):
[11] Nilsson A, Lagesson V, Bornehag CG, Sundell J, Tagesson C. Quantitative
Whether novel biomarkers provide useful information for dis- determination of volatile organic compounds in indoor dust using gas
ease diagnosing, monitoring or risk prediction has been the focus of chromatography-UV spectrometry. Environ Int 2005;31(8):1141–8.
[12] Nilsson A, Kihlström E, Lagesson V, Wessén B, Szponar B, Larsson L, Tagesson C.
intense study [18–20]. Proving the usefulness of such new biomark- Microorganisms and volatile organic compounds in airborne dust from damp
ers, however, has been hampered by a variety of factors including residences. Indoor Air 2004;14(2):74–82.
inadequate statistical power, the lack of measures such as calibra- [13] Lagesson V, Lagesson-Andrasko L, Andrasko J, Baco F. Identification of
compounds and specific functional groups in the wavelength region
tion and reclassification and the lack of external validation [1,2].
168–330 nm using gas chromatography with UV detection. J Chromatogr A
Breathomics might be the new tool that provides the means 2000;867(1–2):187–206.
to differentiate on a physiological basis between clinically related [14] Nelson N, Lagesson V, Nosratabadi AR, Ludvigsson J, Tagesson C. Exhaled iso-
disease profiles or sub-stages. Interestingly, it has already been prene and acetone in newborn infants and in children with diabetes mellitus.
Pediatr Res 1998;44(3):363–7.
suggested that exhaled air analysis might be extra valuable in clin- [15] Lagesson-Andrasko L, Lagesson V, Andrasko J. The use of gas-phase UV spectra
ical practice due to its superior accuracy compared to the reported in the 168–330-nm wavelength region for analytical purposes, 1. Qualitative
sensitivity and specificity of current methodologies [4]. measurements. Anal Chem 1998;70(5):819–26.
[16] Leanderson P, Lagesson V, Tagesson C. Demonstration of nitric oxide on
In conclusion, it can be stated that breath analysis using GC- asbestos and silicon carbide fibers with a new ultraviolet spectrophotometric
DUV holds the promise to be of major interest in clinical practice. assay. Environ Health Perspect 1997;105(Suppl. 5):1037–40.
It has already been proven that in some disease profiles, [17] Jones AW, Lagesson V, Tagesson C. Determination of isoprene in human breath
by thermal desorption gas chromatography with ultraviolet detection. J Chro-
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sensitive and non-invasive methodology [1,2]. Also, GC-DUV may [18] Kubán ˇ P, Foret F. Exhaled breath condensate: determination of non-volatile
be applied to monitor the effects of acupuncture and related compounds and their potential for clinical diagnosis and monitoring. Anal Chim
Acta 2013;805:1–18.
techniques without interfering with the treatment itself.
[19] Sethi S, Nanda R, Chakraborty T. Clinical application of volatile organic
compound analysis for detecting infectious diseases. Clin Microbiol Rev
[20] Kataoka H, Saito K, Kato H, Masuda K. Noninvasive analysis of volatile
biomark-ers in human emanations for health and early disease diagnosis.
[1] Boots AW, van Berkel JJ, Dallinga JW, Smolinska A, Wouters EF, van Schooten Bioanalysis
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and disease. J Breath Res 2012;6(2):027108.

Fig. 3. (a) Example of chromatogram, at wavelength 191 nm, obtained by GC-DUV. (b) Spectrum (_: 160–330 nm) of peek eluting at 13.694 min, obtained by GC-DUV.

that the extraction is fast and simple and that detection limits can the compounds are introduced into the DUV-spectrophotometer
reach parts per trillion (ppt) levels for some compounds. to identify the separated VOCs.
In UV spectroscopy, the sample is irradiated with the broad
2.3. Sample analysis technique spectrum of the UV radiation. If a particular electronic transition
matches the energy of a certain band of UV, it will be absorbed.
A methodology to accurately measure trace gases in complex The remaining UV light (residual radiation) passes through the
mixtures such as exhaled air is based on gas chromatography sample. From this residual radiation a spectrum is obtained, Fig.
anddeep ultra violet spectroscopy (GC-DUV) developed by Chroma- 2a–c, with “gaps” at these discrete energies – which is called an
lytica AB, Malmoe, Sweden. In this method, schematically depicted absorption spectrum.
in Fig. 1, the sample is injected in a GC that separates the dif- GC-UV is as a sensitive method both concerning detection
ferent compounds based on physio-chemical interaction between limits, identification limits and classification limits [10–17]. The
the mobile and column phase. After elution through the column, method is particularly suitable for the identification of structural