Rev Environ Health 2017; 32(1-2): 193–200

Daniela Pelclova*, Vladimir Zdimal, Petr Kacer, Martin Komarc, Zdenka Fenclova,
Stepanka Vlckova, Nadezda Zikova, Jaroslav Schwarz, Otakar Makes, Tomas Navratil,
Sergey Zakharov and Dhimiter Bello

Markers of lipid oxidative damage among office

workers exposed intermittently to air pollutants
including nanoTiO2 particles
DOI 10.1515/reveh-2016-0030 (nano)TiO2 aerosol during their daily visits of the produc-
Received July 28, 2016; accepted September 13, 2016; previously tion area for an average of 14 ± 9 min/day. Median parti-
­published online October 18, 2016 cle number concentration in office workers while in the
production area was 2.32 × 104/cm3. About 80% of the par-
Abstract: Nanoscale titanium dioxide (nanoTiO2) is a
ticles were < 100  nm in diameter. A panel of biomarkers
commercially important nanomaterial used in numerous
of lipid oxidation, specifically malondialdehyde (MDA),
applications. Experimental studies with nanotitania have
4-hydroxy-trans-hexenal (HHE), 4-hydroxy-trans-nonenal
documented lung injury and inflammation, oxidative
(HNE), 8-isoprostaglandin F2α (8-isoprostane), and alde-
stress, and genotoxicity. Production workers in TiO2 man-
hydes C6−C12, were studied in the EBC and urine of office
ufacturing with a high proportion of nanoparticles and a
workers and 14 unexposed controls. Nine markers of lipid
mixture of other air pollutants, such as gases and organic
oxidation were elevated in the EBC of office employees
aerosols, had increased markers of oxidative stress, includ-
relative to controls (p < 0.05); only 8-isoprostane and C11
ing DNA and protein damage, as well as lipid peroxida-
were not increased. Significant association was found in
tion in their exhaled breath condensate (EBC) compared
the multivariate analysis between their employment in
to unexposed controls. Office workers were observed to
the TiO2 production plant and EBC markers of lipid oxida-
get intermittent exposures to nanoTiO2 during their pro-
tion. No association was seen with age, lifestyle factors,
cess monitoring. The aim of this study was to investigate
or environmental air contamination. The EBC markers in
the impact of such short-term exposures on the markers
office employees reached about 50% of the levels meas-
of health effects in office workers relative to production
ured in production workers, and the difference between
workers from the same factory. Twenty-two office employ-
production workers and office employees was highly
ees were examined. They were occupationally exposed to
significant (p < 0.001). None of these biomarkers were
elevated in urine. The approach presented here seems to
be very sensitive and useful for non-invasive monitoring
*Corresponding author: Daniela Pelclova, Department of
of employees exposed to air pollutants, including gases,
Occupational Medicine, First Faculty of Medicine, Charles University
in Prague and General University Hospital in Prague, Na Bojisti 1, organic aerosols, and nanoTiO2, and may prove useful for
120 00 Prague, Czech Republic, E-mail: daniela@pelclova.cz routine biomonitoring purposes. Among them, aldehydes
Vladimir Zdimal, Nadezda Zikova, Jaroslav Schwarz and Otakar C6, C8, C9, and C10 appear to be the most sensitive markers of
Makes: Institute of Chemical Process Fundamentals of the CAS CR, lipid oxidation in similar occupational cohorts. One major
v.v.i., Prague, Czech Republic
challenge with sensitive biomonitoring techniques, how-
Petr Kacer: Institute of Chemical Technology Prague, Czech Republic
Martin Komarc: First Faculty of Medicine, Institute of Informatics,
ever, is their non-specificity and difficulty in interpreting
Charles University in Prague and General University Hospital the meaning of their physiological values in the context of
in Prague, Prague, Czech Republic; and Department of chronic disease development and damage-repair kinetics.
Kinanthropology and Humanities, Faculty of Physical Education and
Sport, Charles University in Prague, Prague, Czech Republic Keywords: exhaled breath condensate; occupational
Zdenka Fenclova, Stepanka Vlckova and Sergey Zakharov: exposure; oxidative stress; spirometry; urine.
Department of Occupational Medicine, First Faculty of Medicine,
Charles University in Prague and General University Hospital in
Prague, Prague, Czech Republic
Tomas Navratil: J. Heyrovský Institute of Physical Chemistry of the
Introduction
CAS CR, v.v.i., Prague, Czech Republic
Dhimiter Bello: Department of Public Health, College of Health The paint and coatings production is known to gen-
Sciences, UMass Lowell, Lowell, MA, USA erate significant particulate matter emissions to the
194      Pelclova et al.: Lipid peroxidation from nanotitania in office workers
atmosphere, including nanoscale particles (1). Titania is
a commonly used wide pigment in paints and coatings.
Unlike bulk particles, titania nanoparticles, that may be
formed during the production, have an enhanced oxidant
capacity and greater potential to induce oxidative stress
and pulmonary inflammation in humans (2). Both experi-
mental and human data show that TiO2 nanoparticles
influence lung physiology; they may exert adverse effects
due to a larger surface area, higher pulmonary deposition
rates, and longer retention as a result of impaired clear-
ance mechanisms and lower detection rates (3). Animals
exposed to TiO2 nanoparticles developed impaired lung
function, pulmonary emphysema, macrophages accumu-
lation, extensive disruption of alveolar septa, and epithe-
lial cell apoptosis (4–6).
In our earlier studies, we utilized the non-invasive
sampling of the exhaled breath condensate (EBC).
During EBC collection, during the tidal breathing into
the condenser, the volatile oxidation products (such as
aldehydes) are exhaled as vapors, whereas the non-vola-
tile compounds are released from the airway lining fluid
in the form of aerosolized particles due to the turbulent
airflow and bubble bursting during opening of the bron-
chioles following exhalation (7). This sampling of the
fluid from the deep airways was combined with a very
sensitive liquid chromatography-electrospray-tandem
mass spectrometry (LC-ESI-MS/MS) analysis of a panel of
oxidative stress markers of nucleic acid damage, protein
oxidation in workers exposed to nanoparticles during
production of white TiO2 and red/brown Fe oxide pig-
ments (8, 9).
In these studies, we have documented elevated
levels of these markers in EBC relative to controls in a
dose-dependent manner. Furthermore, we have also
found increased levels of lipid oxidation markers in pro-
duction workers from the same factory producing TiO2
pigment (10).
As part of earlier studies, we conducted extensive
exposure characterization in the manufacturing plant,
which included generation of area exposure maps in
­different departments, as well as number concentration,
particle size distribution, and mass concentration over a
wide range of particle sizes (10 nm–20 µm) measured by
a scanning mobility particle sizer (SMPS) and an aerody-
namic particle sizer (APS) (both of TSI Inc., MN, USA). The
chemical analysis of the airborne aerosol (fumes and gases
potentially emitted from furnaces) could not be performed.
In the production workshop, the median total mass TiO2
concentration was 0.40  mg/m3. The median number of
concentrations was 23,200 particles/cm3. Importantly,
about 80% of those particles were smaller than 100 nm in
diameter (8).
During the course of the original study, we observed
that office workers would visit the production floor for a
short period of the shift to inspect production processes.
Office workers were included in the original study design
and were studied using identical non-invasive methods
(EBC, urine), workplace observations, and medical exami-
nations. The objective of this work was to investigate in
more detail the impact of intermittent exposures to TiO2 at
the level of biomolecular markers of lipid peroxidation in
the EBC and urine of office workers, relative to unexposed
controls and production workers.
Methods
Subject recruitment
Twenty-two male office employees working in the same building,
where TiO2 pigment was produced, or an adjacent building were
examined. In addition, 14 male control subjects not employed in the
factory were also enrolled. They worked as healthcare personnel and
technical staff and did not handle nanomaterials, including nano-
TiO2, or dusts/aerosols. This later group represented an unexposed
control group.
All participants were interviewed by trained physicians using
a standardized questionnaire concerning personal and occupational
history, medical treatments, and lifestyle habits (diet, alcohol intake,
smoking, and physical activity). Then they underwent a physical
examination, which was followed by the collection of their EBC and
urine. Finally, spirometry was measured. The study was carried out
according to the Helsinki Declaration. The Ethical Committee of the
Charles University approved the study and all participants signed an
informed consent.
Non-invasive sampling and pulmonary function testing
Pulmonary function testing was performed by a SpiroPro, Jaeger,
Germany. The measurements were carried out according to stand-
ard protocols of the German Respiratory Society Guidelines (11). The
measurement included forced vital capacity (FVC), inspiratory vital
capacity (VCIN), peak expiratory flow (PEF), forced expiratory vol-
ume (FEV), and FEV in 1 s (FEV1). FEV1 and FVC were used to meas-
ure the FEV1/FVC ratio.
The post-shift EBC samples were collected at the production
facility using the Jaeger Ecoscreen Turbo DECCS, Jaeger, Germany,
equipped with a filter. All subjects breathed tidally for 15 min through
a mouthpiece connected to the condenser (−20 °C) while wearing a
nose-clip; the collection was performed in accordance with the
guidelines of the American Thoracic Society and European Respira-
tory Society (7). All EBC and urine samples were immediately frozen
and stored at −80 °C until analysis within 2 months.
 Pelclova et al.: Lipid peroxidation from nanotitania in office workers      195

Oxidation products of malondialdehyde (MDA), 4-hydroxy-trans-

hexenal (HHE), 4-hydroxy-trans-nonenal (HNE), 8-isoprostaglandin F2α
(8-isoprostane), and C6–C12 hydrocarbons (aldehydes) were analyzed in
the EBC and urine as was previously described (12, 13). Briefly, samples
were purified and concentrated by solid-phase extraction (SPE) followed
by LC-ESI-MS/MS analysis. Deuterium-labeled internal standards were
added to the samples prior to SPE as described in the above-referenced
papers. To exclude contamination of EBC by saliva, the concentration of
α-amylase was also monitored as previously described (14). The concen-
tration of urinary biomarkers was adjusted for creatinine concentration
to correct for variations in urine dilution rates (15).
Environmental pollution
Information on major environmental air pollutants, SO2, NOx, and O3,
was collected from local monitoring stations for the days and loca-
tions of the EBC sampling to assess the potential confounding effect
on oxidative stress markers (16).
Statistical analysis
Results
Subjects
The mean age of 22 office workers did not differ signifi-
cantly from the age of 14 controls (44.3 ± 3.9  years and
38.5 ± 4.5 years, respectively). The lifestyle habits, includ-
ing alcohol consumption and physical activity, were com-
parable in both groups. Only the proportion of smokers in
the group of office employees (4.5%) was lower (p < 0.01)
than that in the controls (50.0%). The two groups were
generally healthy, only asthma was diagnosed in two
(9.1%) subjects from the office group and in one control
subject (7.1%). Chronic bronchitis symptoms were present
in three males in the control group only (21.4%). The
mean values of the pulmonary function did not differ
between the two groups of subjects and were within
normal ranges.

Statistical analysis included test for differences between the two

groups under study (office workers and unexposed controls) using
χ2  test, t-test, or Mann-Whitney test. Correlation and regression
­analyses were used to assess the relationship between the variables
of interest. All analyses were conducted using SPSS V.22.0 (SPSS,
Inc., Chicago, Illinois, USA).
Exposures
The length of office employee’s employment in the
factory was 15.5 ± 3.6 years. Their offices were located at

Figure 1: Markers of oxidation of lipids in exhaled breath condensate.

Mean and 95% confidence interval (CI) are shown. Malondialdehyde (MDA), 8-isoprostaglandin F2α (8-isoprostane), 4-hydroxy-trans-hexe-
nal (HHE), 4-hydroxy-trans-nonenal (HNE), and aldehydes C6–C10. ***p < 0.001.
196      Pelclova et al.: Lipid peroxidation from nanotitania in office workers

0.02
(−0.02, 0.06)
0.00
(0.00, 0.00)

0.00
(0.00, 0.00)
0.00
(0.00, 0.00)
C11

0.06b
(0.02, 0.10)

0.00a
(−0.01, 0.00)
0.00b
(0.00, 0.00)
Table 1: Multiple regression analysis (regression coefficient and 95% CI) of the elevated lipid oxidation markers in the exhaled breath condensate of administrative/office workers with various

−0.01 
0.00 

0.00 

0.00 
C10 

1.17  

−0.03 

−0.01 
C

(−0.44, 0.43)
(−0.02, 0.02)

(−0.01, 0.01)

(−0.01, 0.01)
(0.71, 1.63)

(−0.07, 0.01)

(−0.01, 0.00)
0.02 
0.01 

0.00 
C 9 

1.48  

−0.02 

−0.01 

0.00 
C

(−0.51, 0.55)
(−0.01, 0.03)

(0.00, 0.01)
(0.92, 2.03)

(−0.06, 0.03)

(−0.01, 0.00)

(−0.01, 0.01)
Figure 2: Markers of oxidation of lipids (aldehydes C11–C12) in
exhaled breath condensate.
Mean and 95% confidence interval (CI) are shown. **p < 0.01.

−0.38 
−0.02 
C8 

1.14  

−0.03 

0.00 
−0.01 

0.00 
C

(−0.97, 0.21)
(−0.04, 0.01)
(0.53, 1.76)

(−0.08, 0.02)

(−0.01, 0.01)
(−0.02, 0.00)

(−0.01, 0.02)
higher levels of the building, where titania pigment was
produced at the ground and first level, or in an adja-

MDA, malondialdehyde; HNE, 4-hydroxy-trans-nonenal; HHE, 4-hydroxy-trans-hexenal. a(p < 0.05), b(p < 0.01), and c(p < 0.001).
cent building of the factory. The office employees visited
the production floor for a daily average of 0.23 ± 0.15  h
(14 ± 9 min). There have been no major changes in the pro-

0.41 
C7 

0.83 

0.00 

0.03 

−0.01 
0.01 

0.01 
(−0.46, 1.29)

(−0.02, 0.01)
(−0.09, 1.75)

(−0.03, 0.04)

(−0.04, 0.10)

(−0.01, 0.02)

(−0.01, 0.03)
duction technology or production volumes over the course
of the past decade. From the area maps of total number
concentration, and previously reported exposure meas-
urements in the production area (8), office workers may
other factors (their age, smoking, body mass index, and environmental air contamination in the city).

be exposed to a ­geometric mean of 25,000 particles/cm3

0.11 

0.00 
C6 

2.07  

0.00 

−0.01 

−0.02a 

0.00 
C

(−0.77, 1.00)

(−0.01, 0.01)
(1.15, 3.00)

(−0.03, 0.03)

(−0.09, 0.06)

(−0.03, 0.00)

(−0.02, 0.02)
(geometric ­standard deviation, 2.2), an arithmetic mean of
27,000 particles/cm3 (maximum of 90,000 particles/cm3),
PM0.1 (­particulate matter < 100 nm) mass concentration of
~5 µg/m3, and PM10, ~3 mg/m3.
−0.60 
HHE 

4.65  

−0.09 

0.07 

0.02 

0.01 

−0.02 
(−4.64, 3.45)
a

(0.42, 8.88)

(−0.24, 0.06)

(−0.27, 0.41)

(−0.04, 0.08)

(−0.05, 0.06)

(−0.12, 0.08)
Biomarkers in EBC and urine

In the EBC of the office employees from the TiO2 producing

plant, the levels of the majority of markers of lipid oxida-
3.87 
HNE 

5.96  

0.01 

−0.13 

−0.03 

−0.05 

0.13 
(−1.55, 9.30)
a

(0.28, 11.64)

(−0.20, 0.21)

(−0.59, 0.33)

(−0.11, 0.06)

(−0.12, 0.02)

(−0.01, 0.26)

tion were higher (p < 0.05) compared to the controls, as

can be seen in Figures 1 and 2. Of all EBC markers, only
8-isoprostane and aldehyde C12 were not significantly
increased. There was no correlation of the markers of
oxidation of lipids with the respiratory or systemic dis-
−0.38 
MDA 

6.89  

−0.14 

−0.15 

−0.03 

−0.01 

0.11 
(−5.63, 4.86)
a

(1.40, 12.38)

(−0.34, 0.06)

(−0.59, 0.30)

(−0.11, 0.05)

(−0.08, 0.05)

(−0.03, 0.24)

orders (hypertension, increased cholesterol, cancer, and

­diabetes) in the office workers and controls.
Multiple regression analysis of occupational expo-
sure, age, smoking, and daily alcohol consumption con-
firmed a significant association between occupational
Smoking (yes/no) 

exposure in the (nano)TiO2 production facility and EBC

 

Administrative/  

Body mass index  

 
factory (yes/no)

level of markers of oxidation of lipids, as shown in Table 1.

SO2 (µg/m3)

NOx (µg/m3)
Age (years)

Urine concentrations of the oxidative stress markers

O3 (µg/m3)
(kg/m2)

were not elevated relative to the control group (data not

shown).
 Pelclova et al.: Lipid peroxidation from nanotitania in office workers      197
The level of air pollution was classified by the
National Hydrometeorological monitoring system as very
low or low (data not shown). No values exceeded the rec-
ommended limits for any of the criteria pollutants, includ-
ing SO2, NOx, and O3.
Discussion
Oxidative stress is a commonly observed mechanism of
particle and nanoparticle toxicity in in vitro and in vivo
systems, including humans. Nanotitania and other types
of nanoparticles, incidental or engineered, exhibit dif-
ferent levels of oxidative stress. NanoTiO2 anatase is a
well-known catalyst and shows low-to-moderate oxida-
tive stress potential in biological systems relative to other
more potent oxidants (such as copper oxide and manga-
nese oxide nanoparticles) (17).
We found elevated (and statistically significant) levels
of oxidative stress markers in the EBC of office workers
employed at the (nano)TiO2 production facility, in compar-
ison with unexposed controls. The levels of EBC biomark-
ers in office workers were roughly half the levels found
in post-shift EBC samples of the production workers and
the differences were statistically significant (p < 0.001)
for all measured markers (10). In contrast, environmental
air pollutants (SO2, NOx, and O3) in the urban air were not
associated with EBC markers. It is reasonable to postulate
that these elevated EBC biomarkers in office workers are
likely caused by repeated short-term exposures, averaging
14 min/day duration, over months and years.
Differences in the levels of the markers of oxidative
stress were also seen earlier among groups of workers
in the different parts of this white pigment production
process and they were related to different levels of aerosol
exposure in our 2-year study (8). We have shown that a
gradient of exposures to nanoTiO2 in different exposure
groups produces measurable (and statistically signifi-
cant) responses in EBC markers. This is the first report
to document increased EBC biomarker levels in response
to repeated exposures to (nano)TiO2 (or any other nano-
material for that matter) of such short-term duration.
These findings speak to the sensitivity of the EBC analysis
coupled with LC-ESI-MS/MS techniques for biomonitoring
purposes.
Interpretation of the biological significance of the
measured biomarker levels in the context of disease pro-
gression is of great interest but challenging. Subjective
respiratory and/or allergy symptoms were not more fre-
quent in the office workers than in the control subjects.
Lung function in our office workers was not impaired,
which is in agreement with the fact that even in our earlier
study of the (nano)TiO2 production workers, all lung
function parameters except two were normal. The only
two parameters that were decreased, i.e. PEF and VCIN,
were found in smoking workers with longer than average
occupational exposure (18). This is in accordance with
the results of the few human studies published to date.
However, experimental studies with higher exposure
doses have resulted in obstructive ventilatory changes,
including impairment of PEF (19) and corresponding
histological changes (19, 20). Similarly, MDA, HHE and
n-hexanal levels were higher in the EBC of nine produc-
tion workers exposed to multi-walled carbon nanotubes
compared to four office workers, while both groups had
normal lung functions (21).
EBC is thought to closely reflect the composition of
the bronchoalveolar lining fluid, and MDA, HNE, HHE,
and 8-isoprostane are known to increase during oxidative
stress. They have been used to monitor lung damage in
patients with asthma and chronic obstructive pulmonary
disease (22, 23), similarly in pneumoconioses caused by
carcinogenic dust asbestos (24, 25), and silica (26). They
were not increased by systemic diseases (27) that may
affect plasma or urine markers (13).
The current study does not allow commenting on or
investigating damage-repair kinetics because only one
measurement/person was available. In an experimental
study, transient changes in the bronchoalveolar lavage
fluid were observed after a single intra-tracheal dose for
1 month; and nanoparticles in the cells were seen still
6 months later (28). In the Lee et al. study of workers han-
dling carbon nanotubes, post-shift MDA, HHE, and n-hex-
anal levels remained elevated until late in the evening
(21). Similarly, we have found that elevated oxidative
stress markers in (nano)TiO2 workers persisted until the
following shift (10), and so did TiO2 particles in EBC (29).
Perhaps the repair mechanisms are sufficient to compen-
sate for the limited lung damage due to oxidative stress
following single exposure episodes in this study. It would
be of great interest for future studies to study in detail
the mechanisms of such damage-repair kinetics using
high density sampling approaches and clearance data.
The impact of current exposures and sustained oxidative
stress in other organ systems, especially cardiovascular,
liver, immunologic and carcinogenesis, and in suscepti-
ble subpopulations of workers, especially smokers, is not
known, but important to study.
Human studies involving engineered nanoparticles
are limited (30). One study found a decrease in antioxidant
enzymes and elevated cardiovascular markers in workers
198      Pelclova et al.: Lipid peroxidation from nanotitania in office workers
handling nanomaterials (31). In a controlled study, healthy
volunteers exposed to nanoparticles from photocopiers
(which are a mixture of incidental nanoparticles gener-
ated during photocopying and engineered nanoparticles
present in toners and paper) exhibited increased oxidative
stress and upper respiratory tract inflammation as well as
oxidative stress in urine of healthy volunteers (32), which
did not clear until 24–36 h post-exposures. These findings
were substantiated in subsequent studies (33–35).
In office workers exposed to nanoTiO2, aldehydes
­C6–C12 were measured for the first time and they proved
even higher elevation than previously mentioned markers.
Linear aldehydes, such as C6, C7, and C9, etc., are the end
metabolites of the lipid peroxidation process of ω-3 and
ω-6 fatty acids, which are principal components of the
phospholipids that form a key portion of cell membranes
(36). Among them, C6, C8, and C9 concentrations in exhaled
air were significantly higher in lung cancer patients than
in smokers and healthy controls (37). It has been hypoth-
esized that exhaled aldehydes reflect both oxidative stress
microenvironments in tissues and tumor-specific tissue
composition and metabolism, and might be used as a non-
invasive probe for lung malignancies (38).
There is no clear explanation why 8-isoprostane and
C12 were not significantly elevated in the office workers.
The reason may be the relatively low number of subjects
in the groups and a higher proportion of smokers in the
control group which might have increased the results in
this group.
The lipid peroxidation data complement our recent
findings of elevated markers of nucleic acid and protein
oxidation in the identical group of office employees (39),
for whom a similar pattern of exposure-biomarker rela-
tionships has been documented.
Limitations
Markers of oxidative stress are not specific for the effect
of the nanoparticles, and other causative factors or con-
founders cannot be completely ruled out. We acknowl-
edge that nanoparticles other than TiO2 may have been
present in some areas of the production plant. Further-
more, we recognize the opportunity for better personal
quantitative exposure assessment. Gaseous pollutants
and other incidental nanoparticles associated with TiO2
production may have contributed to oxidative stress in
production and office workers. The chemical analysis of
the airborne aerosol on filters could not be performed due
to the limited resources and limit of detection issues. It
would also have been highly desirable to monitor for other
co-exposures in the production workshops, such as fumes
and gases potentially emitted from furnaces. However,
diesel exhaust in the transport corridors did not play an
important role as the real-time aerosol exposures in these
locations were low.
Due to limited resources, aerosol concentrations were
not measured in the rooms of the office employees from
the factory. As also the control group spent most of the
time in an office room with comparable technical equip-
ment, we suppose that the visit of the production area of
the factory was the key factor.
A major problem is the absence of physiological refer-
ence values for lipid oxidation markers in EBC, and it is
possible that EBC markers in the office employees may be
situated at the upper level of the normal reference range.
EBC collection and examination is not yet a standard-
ized method, and it may be influenced by the condensing
equipment and other factors. To address this, a control
group must always be examined in parallel, which was
also included in our study (7).
Conclusion
Nanoparticle products are becoming more and more prev-
alent in the marketplace and in our environment, and their
potential harm to humans, especially workers, should be
the focus of intense research. Our results are in agreement
with experimental studies and with our and other studies
in workers and volunteers exposed to nano-sized parti-
cles and a mixture of air pollutants, including gases and
organic aerosols, all causing oxidative damage (8, 18, 22).
Levels of lipid ­oxidation markers in the EBC exhibited an
exposure-response gradient: unexposed controls < office
workers (15  min/day) < production workers (8 h/day, 5
days/week). The differences between exposure groups
suggest that lowering exposure may help to decrease this
biological effect within time, although the kinetics of such
damage repair is unknown.
Lung function measurement does not appear to
be a sensitive enough technique for detection of early
changes, but it is important in assessing the decline in
lung function. Non-invasive measurement of appropri-
ate biomolecular markers reflecting molecular injury
pathways, such as oxidative stress, DNA, and protein
damage, etc., in the EBC seems to be a highly sensitive
technique and a method of choice to monitor early bio-
molecular effects of exposure to nanoparticles in work-
places. In addition, it enables measurement of particles
(20), metals (18), markers of oxidation of nucleic acids
 Pelclova et al.: Lipid peroxidation from nanotitania in office workers      199
and proteins (8), and markers of inflammation (18) in
the EBC.
More attention should be paid to both production
and office employees in the factories with nanoparticle
exposure, and a more detailed chemical analysis of their
workplace aerosol is needed. Our experience is that a
combination of sensitive non-invasive techniques, such as
EBC–LC-ESI-MS/MS, complemented with tests that assess
the capacity of an organ’s function (spirometry) provides
a more comprehensive picture of the damage and possible
disease development in workers.
Acknowledgments: The authors would like to thank
the Charles University in Prague projects P25/1LF/2,
P28/1LF/6, and EU Project, Material and technical base
for the research of diagnostics and therapy of civilization
and oncological diseases and their significant risk factors
in the General University Hospital in Prague” (reg. No.
CZ.2.16/3.1.00/24012).
Conflict of interests: The authors claim no competing
financial or non-financial conflicts of interests.
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