〈一般研究課題〉 三次喫煙評価のための微量分析法の開発

助 成 研 究 者

豊橋技術科学大学

齊戸美弘

三次喫煙評価のための微量分析法の開発
齊戸 美弘（豊橋技術科学大学）

Determination of Volatile Organic Compounds for a Systematic
Evaluation of Third-Hand Smoking
Yoshihiro Saito（Toyohashi University of Technology）

Abstract:
Third-hand smoking was quantitatively evaluated with a polymer-packed sample preparationneedle and subsequent gas chromatography-mass spectroscopy（GC-MS）analysis. The extractionneedle was prepared with polymeric particles as the extraction medium, and successful extractionof typical gaseous volatile organic compounds（VOCs）was accomplished with the extraction needle.
For an evaluation of this new cigarette hazard, several types of clothing fabrics were exposed tosidestream smoke, and the smoking-related VOCs evaporated from the fabrics to the environmental
air were preconcentrated with the extraction needle. Smoking-related VOCs in smokers' breath
were also measured using the extraction needle, and the eﬀect of the breath VOCs on third-handsmoking pollution was evaluated.
The results demonstrated that a trace amount of smoking - related VOCs was successfully
determined by the proposed method. The adsorption and desorption behaviors of smoking-related
VOCs were clearly different for each fabric material, and the time variations of these VOCs
concentrations were quantitatively evaluated. The VOCs in the smokers' breath were clearly higher
than that of nonsmokers'; however, the results suggested that no signiﬁcant eﬀect of the smokers'
breath on the potential pollution occurred in the typical life space. The method was further applied
to the determination of the actual third-hand smoking pollution in an automobile, and a future
possibility of the proposed method to the analysis of trace amounts of VOCs in environmental air
samples was suggested.

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1. Introduction
It has been reported that tobacco smoke contains more than 4000 chemicals containing potential
carcinogens［1,2］. The analysis of environmental tobacco smoke（ETS）has been focused on for asystematic evaluation of indoor air pollution, because the evaluation of passive smoking could be
important to avoid any risk to nonsmokers' health. In recent years, the expose to passive smoking
risk has been reduced by adopting smoke-free areas or smoking rooms; however, a new smoking
hazard has been reported, so-called "third-hand smoking"［3］.
Third-hand smoking is residual tobacco smoke pollution, that occurs after smoking, and isclassiﬁed into the following two types: one is in-door air pollution due to tobacco-related volatileorganic compounds（VOCs）from in-door surfaces, such as cushions, carpets or curtains. These
VOCs are adsorbed on in-door surfaces during smoking, and are then desorbed form the surfaces
after smoking, even if the smoke has already been ventilated. Another is exposure to tobaccorelated VOCs from a smoker. Although smoking is done outside, tobacco-related VOCs still remainon the surface of the smoker's clothing and also the smoker's breath. Some of these chemicalcompounds have an unpleasant smell for nonsmokers, and potential health damage has beensuspected, especially for children［3］.
Gas chromatography（GC）is one of the most promising methods for the analysis of smokingrelated VOCs, because of their selectivity, and gas chromatography-mass spectrometry（GC-MS）
has been realized as a powerful tool for a sensitive determination of typical VOCs and semi-volatileorganic compounds（SVOCs）in tobacco smoke［4,5］. However, a sample preconcentration process
should still be needed for accurate and precise determination of these VOCs. Especially, the VOCs
concentration related to third-hand smoking could be at a quite low level, and therefore an eﬀective
sample preparation process and subsequent separation/determination methods are needed.
Solid-phase microextraction（SPME）is one of the most advanced sampling techniques fortypical VOCs analysis in GC［6］, and it has several attractive features over traditional sample
preparation techniques. Alternative sample preparation techniques have been developed as a needletype sample preparation device［7-15］. Compared to the conventional SPME, the needle-type sampling
device has been regarded to have good mechanical strength and high extraction capacity. In ourprevious work, a polymer particulate medium based on a copolymer of methacrylic acid（MA）and
ethylene glycol dimethacrylate（EDMA）was synthesized, and packed into a specially-designed
needle as the extraction medium［9］. This polymer particle - packed extraction needle was
successfully applied to the analysis of ETS［11］and human breath［16］.
In this paper, a polymer particle-packed extraction needle was applied to the evaluation of apotential risk of third-hand smoking. Smoking-related VOCs adsorbed on several types of fabrics
were determined, and the time variation of the VOCs concentration was studied. Smoking-related
VOCs in smokers' breath were also measured using the extraction needle, and the eﬀect of thebreath VOCs on the third-hand smoking pollution was evaluated. Additionally, the proposed method
was applied to the determination of actual third-hand smoking in an automobile.

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2. Experimental

Benzene, toluene, xylenes and 2 , 5 -dimethylfurane（2 , 5 -DMF）were obtained from Tokyo
Chemical Industries（Tokyo, Japan）. Acetone, pyrrole, limonene and 2 -furaldehyde werepurchased from Wako Pure Chemical（Osaka, Japan）. All of the fabric samples（cotton, linen,
silk, acetate and polyester）used in this work were commercially available, and purchased from a
local fabric store（Toyohashi, Japan）. Breath samples were collected in Tedlar bags（GLSciences, Tokyo, Japan）. A vacuum sampling device and an ammonia gas detection tube werepurchased from Komyo Rikagaku Kogyo（Tokyo, Japan）.

As the extraction medium, a copolymer of MA and EDMA was synthesized as described
previously［9］, and copolymer beads having a diameter of between 150 and 180 μm were packed
into a section of a specially-prepared needle, 0.5 mm i.d., 0.7 mm o.d., 85 mm length. For the
extraction, the needle was attached to a commercially available vacuum sampling device. A
gaseous sample was introduced into the extraction needle by manually pulling the handle of thevacuum sampling device, and the analytes were extracted by the polymeric particles therein. The
typical sampling volume was 50 mL, and it took about 5 min to complete the sampling.
After extraction, the extraction needle was attached to an injection syringe containing 0.5 mLof N 2 gas. Then, the extraction needle was inserted to a heated GC injection port, and theanalytes were injected by the N2 gas, where the needle was heated for 10 s in the injector before
injection. It has been conﬁrmed in preliminary experiments that the pre-heating time could help
realize successful desorption of the analytes. The extracted samples could be stored in theneedle at room temperature for at least three days.

In this work, the risk of third-hand smoking was evaluated based on the determination ofsmoking-related VOCs that adsorbed on several types of fabrics, and that were also contained in
the smokers' breath. Figure 1 shows the scheme for the evaluation of smoking-related VOCs
desorbed from fabric samples. First, a 10 cm square of fabric sample was placed in a home-mademetal box of 18 L volume. Then, tobacco was placed on the bottom of the box. The box had an
inlet, as illustrated in Fig. 1 , where enough air had been provided during tobacco combustion.
During tobacco combustion, the box was ﬁlled with sidestream smoke and smoking-related VOCs
were adsorbed on the fabric. After 9 min of combustion time, the fabric was transferred to a
glass vessel of 4 L volume. Upon completion of the equilibrium time for 30 min in the glass
vessel, the extraction needle was inserted into the glass vessel, and then 50 mL of the gaseous
sample was extracted, followed by GC-MS analysis. One of the unpleasant odors, ammonia, wasmeasured by using a commercially-available detection tube.

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Breath samples of smokers and nonsmokers were collected in a Tedlar bag of 1 L volume, andthen extracted with the extraction needle. Tedlar bags were cleaned at least ﬁve times with pureN2 before use.
Extraction needle
10 cm X 10 cm
clothing fabricBurning
tobacco18 L metal box

4 L glass vessel

Figure 1 Scheme for the evaluation of smoking-related VOCs level desorbed from fabrics withneedle extraction device.

For all of the GC measurements, an Agilent 6890N gas chromatograph (Yokogawa Analytical
Systems, Musashino, Japan) with a split/splitless injection port and a HP 5972A mass selective
detector (MSD) was used, and all of the injections were made by a split mode with a ratio of
15:1. GC separation was performed on an HR- 17 fused-silica capillary column of 30 m x 0.25
mm i.d. having a ﬁlm thickness of 0.25 μm (Shinwa Chemical Industries, Kyoto, Japan). As thecarrier gas, He was employed at the typical head pressure of the 30 kPa, and the injector wasmaintained at 200℃. The GC-MS interface temperature and the ionization voltage were set at
250 ℃ and 70 eV, respectively, with electron impact ionization. The column temperature wasstarted at 40 ℃ for 2. 0 min, and then programmed to 240 ℃ at a rate of 10 ℃/min. The mass
spectrometer was operated in either a total ion monitoring mode (TIM) with a m/z range of 45 to250, or a selected ion-monitoring mode (SIM). The data collection was made with Chrom NAV(Jasco, Tokyo, Japan) running on a personal computer.

3. Results and Discussion

As an evaluation method for the air pollution generated by the third-hand smoking, thedetermination of VOCs desorbed from various types of fabrics could be quite valuable. Several
experimental conditions, such as the volume of the metal box and the glass desorption vessel, thesize of the fabric, and the equilibrium time of tobacco-related VOCs in the glass vessel, were
optimized in preliminary experiments to accomplish a reproducible and eﬀective extraction. For
all of the evaluations, a 10 cm square of each fabric sample was used, because the physical sizeof the fabric showed a dominant eﬀect on the amount of the VOCs than that of the weight in the
preliminary experiments. This is quite natural that the amounts of desorbed VOCs are more

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closely related to the surface area of the fabric samples than the weight [17], and actually thesame size of fabric sample should be examined for a more practical evaluation of third-handsmoking in an in-room environment.
Figure 2A shows typical TIM chromatogram for the separation of VOCs related to third-handsmoking. Introducing the TIM mode, a large number of VOCs related to a third-hand smoking
have been successfully detected in GC-MS with the sample preconcentration by the extractionneedle, although the detected VOCs level was clearly lower than that detected in typicalsidestream smoke. In addition, a similar VOCs proﬁle was obtained for several diﬀerent brandsof cigarettes, except for that containing a menthol ﬂavor, where the peak of menthol was clearlyobserved in the chromatogram. Also, in the sidestream smoke, the peak of nicotine was detected;
however, it was not detected in samples related to the third-hand smoking. The above results
have a good agreement with phenomena reported previously, where a quite limited desorption,
due to the low volatility, of nicotine from the surface of the fabrics was conﬁrmed [18]. For asensitive determination of VOCs closely related to third-hand smoking, pyrrole, benzene, tolueneand 2,5 -DMF were chosen as the target compounds, and monitored with a SIM mode of m/z 67,
78, 91 and 96, respectively. A typical SIM chromatogram for the separation of VOCs related tothird-hand smoking is summarized in Fig. 2B. The chromatogram was constructed from the sumof these m/zs' responses. The limit of detection (LOD) and the limit of quantiﬁcation (LOQ) for
all these compounds are summarized in Table 1. The RSDs of the peak area were less than 3.0%
(n= 5 ) for all of the standard samples. The extraction/desorption efficiency of the extractionneedle was evaluated by comparing the corresponding peak areas for the direct injection of thehigher concentration VOCs sample and needle extraction of the lower concentration sample.
From this comparison, the sample recovery with the extraction needle was calculated as being
more than 99% in the calibration range. The upper limits of the calibration range were more than500

μ g/L for all VOCs, suggesting that the extraction needle has a satisfactory extraction

performance for these VOCs.
(A)

(B)
f

a

MS abundance / -MS abundance / -

f

d
b

c

d

c
e

e

2

4

6

a
8

10

12

2

b
4

6

8

10

12

Retention time / min

Retention time / minFigure 2 Typical chromatograms of the smoking-related VOCs adsorbed on an acetate fabric.（A）Monitored by TIM mode and（B）monitored by SIM mode. Peaks:（a）benzene;（b）2 , 5 -DMF;（c）toluene;（d）pyrrole;（e）-xylene; and（f）2 -furaldehyde.

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Table 1 LOD and LOQ for four typical smoking-related VOCs.

Benzene2,5 -DMF

ToluenePyrroleLOD（ng/L）

1.0

1.0

0.5

5.0

LOQ（ng/L）

3.0

3.0

1.5

15

The chromatograms were monitored as the sum of the responses atfour m/z's : 67, 78, 91 and 96.

Five different types of fabrics (cotton, linen, silk, acetate and polyester) were exposed tosidestream smoke, and the VOCs related to the third-hand smoking were determined. Figure 3
presents the concentration of the smoking-related VOCs desorbed from the fabric samples in theglass vessel. Signiﬁcantly higher concentrations for ammonia and 2 -furaldehyde (data not shown)
were observed for the cotton and linen fabric samples; however, higher concentrations for othersmoking-related aromatic VOCs were obtained with the acetate fabric. The results show good

-1

6
4
2

60

Toluene

50
40
30
20
10
0

12
8
4

1.0

Cotton
Silk
PolyesterLinen
AcetatePyrrole

0.8
0.6
0.4
0.2
0

Cotton
Silk
PolyesterLinen
AcetateBenzene16

0

Cotton
Silk
PolyesterLinen
Acetate

-1

-1

20

Concentration / ng L

80

Concentration / ng L

Ammonia

Concentration / μ g L

Concentration / μ g L-1

10

Cotton
Silk
PolyesterLinen
AcetateFigure 3 Comparison of smoking-related VOCs adsorbed on each fabric. Concentration of ammonia
was determined by a commercially available detection tube. Other compounds were determined by
GC-MS in the SIM mode.

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agreement with that reported previously [17], where a quite limited adsorption of smoking-related
VOCs was observed for the polyester and silk fabric samples. Although a more extensive study,
including a consideration for the chemical structure of the ﬁber, should be needed to reach theﬁnal conclusion, these results indicate that the adsorption behavior of VOCs was associated withthe chemical structure of the fabric, and that the proposed method could be applied to thedevelopment of an evaluation method for preventing third-hand smoking pollution.
In an indoor environment, tobacco-related VOCs are rapidly diffused to indoor air, andadsorbed on various indoor surfaces. The adsorbed VOCs are re-emitted to the indoor air
gradually. Therefore, a time-variation profile of the VOCs evaporated from the fabric wasdetermined for an evaluation of third-hand smoking pollution. After the smoking-related VOCs
were adsorbed on a fabric in the metal box, the fabric was placed in calm outdoor air for 1 to 10
min. During this period, the adsorbed VOCs were progressively evaporated from the surface ofthe fabric to ambient air. Then, the fabric was transferred to a glass vessel of 4 L volume, andthe VOCs in the vessel were extracted with the extraction needle after 30 min of theequilibration time. The time variations of the typical VOCs level are shown in Fig. 4, where theconcentration is plotted against the exposed time of the fabric to the outdoor air. It can be seenthat these VOCs were continuously evaporated from the fabric surface for about 10 min, andalso that the compounds having high volatility were more rapidly evaporated from the fabric.

Ammonia

Benzene4

Concentration / μg L

-1Concentration / ng L-1

16

3
2

1
0

0

2

4

6

8

12
8

4

0

10

0

2

Time / min
Toluene1.0

50Concentration / μ g L-1

Concentration / ng L

-1

60

40
30
20
10
0

0

2

4

6

4

6

8

10

8

10

Time / min

8

10Time / min

Pyrrole

0.8
0.6
0.4
0.2
0

0

2

4

6

Time / minFigure 4 Time variation profile of the concentration of smoking-related VOCs adsorbed on acetate fabric.

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A large number of tobacco-related VOCs are included in the smokers' breath [19], and thesecompounds could be one of the origins of third - hand smoking. Figure 5 shows a typicalchromatogram of a smoker's breath sample. The breath sample was collected at just after theﬁnal puﬀ. The peaks of acetone, benzene and toluene were also detected in nonsmoker's breath
samples; however, these concentrations were clearly higher in the smoker's breath. Additionally,
many VOCs were only detected in the smokers' breath; especially, 2,5 -DMF has been recognized
as a tobacco smoke biomarker in the smoker's breath [19,20], and actually detected only in thesmokers' breath in this work. For all of the smokers' breath samples, the level of ammonia was
below the LOD of the commercially-available detection tube.

MS abundance / -

d

f

a

b c

2

4

e

6

8

10

12

Retention time / minFigure 5 Typical chromatogram of the tobacco-related VOCs in smoker's breath. Peaks:（a）acetone;（b）benzene;（c）2 , 5 -DMF;（d）toluene;（e）o, m-xylene; and（f）limonene.

The time variations of the tobacco-related VOCs level in smoker's breath are illustrated inFig. 6, where the concentrations of these VOCs are plotted against the elapsed time from theﬁnal puﬀ of the cigarette. It was conﬁrmed that tobacco-related VOCs were continuously expired
from the smoker's breath for about 10 min, although the concentrations of these VOCs weresigniﬁcantly higher over the ﬁrst 5 min.
Using a model case, the eﬀect of the smoker's breath on the third-hand smoking pollution was
evaluated. The amount of the expired benzene from the smoker's breath was calculated as 280 ng
by the sum of the expired benzene during a period of 10 min, from 0 . 25 to 10 . 25 min aftersmoking, where the typical tidal volume and respiratory rate of healthy adults of 500 mL and 15
times/min, respectively, were used. If a smoker enters into a typical non-smoking room of 22.5
m3 , 3 m x 3 m x 2.5 m, after smoking outside or in a smoking-room, the concentration of benzene
could be estimated as 0.012 ng/L in the in-room environment. The above results suggest that any
signiﬁcant contribution of the smokers' breath to the concentrations of smoking-related VOCs in
the room was not confirmed by this model. However, tobacco-related VOCs were initiallyexhausted from the smokers' breath at a relatively high level, and a potential health hazard fromthe third-hand smoking hazard could be suspected if contact with a smoker immediately after− 104 −

Benzene

20
15
10
5
0

0

2

4

6

Time / min

8

15
10
5

0

55

Concentration / ng L-1

-1
30

20
10

2

4

6

2

4

6

8

10

8

10

Time / min40

Concentration / ng L

20

0

10

Toluene00

2,5-DMF

25

Concentration / ng L-1

Concentration / ng L

-1

25

8

Limonene

50
45
40
35
30
25
20
0

10

Time / min

2

4

6

Time / minFigure 6 Time variation profiles of the concentration of tobacco-related VOCs in smoker's breath.

his/her smoking. The total amount of the smoking-related VOCs exhausted form a smoker was
clearly lower than that in a typical mainstream smoke, suggesting that most of the VOCs in themainstream smoke were absorbed into the body from the lungs, and only a part of these VOCs
were expired as breath.

As described previously, the extraction needle showed a good reproducibility and storage
performance for typical VOCs [9], and these features indicated the suitability of the extractionneedle to real outdoor sample analysis. The applicability of the present method to actual thirdhand smoking evaluation was carried out based on the determination of smoking-related VOCs in
a compact automobile having an interior volume of 3 . 3 m 3 . After measurements of the VOCs
concentrations, two cigarettes were burned in a car for a period of 9 min, while all the windows
and doors were closed. After combustion, these cigarettes were removed from the car andsecond-hand smoke was maintained for 5 min in the car. Then, all of the doors were opened for 1
min to remove the second - hand smoke from the car completely. Upon completion of air− 105 −

Concentration / ng L

-1

10

8

XyleneTolueneBenzene2,5-DMF

6

4

2

0

10

20

30

40

50

Time / minFigure 7 Determination of actual third-hand smoking pollution in a car. The concentration of xylene
was determined as m-xylene.

ventilation, all of the doors were closed again, allowing the smoking-related VOCs adsorbed on
the indoor surface to re-emit into the air, although a part of the adsorbed VOCs could alreadybe desorbed during the ventilation process. The time variations of these VOCs were monitored
using the extraction needle as the sample preparation device. As can be found in Fig. 7 , theconcentrations of tobacco-related VOCs were gradually increased after closing the doors,suggesting a slow desorption of tobacco-related VOCs from the car interior surface. In thismodel study, the compounds having a relatively higher volatility, such as benzene and 2,5 -DMF,
showed an equilibrium concentration within 10 min; however, compounds with a lower volatility,
such as toluene and xylenes, needed about 20 min to reach the plateau. Although a morecomprehensive study should be scheduled to precisely evaluate the real risk of third-handsmoking, the above results demonstrated a successful application of the extraction needle as asampling/preconcentration device for the analysis of smoking-related VOCs in air samples, andalso suggest the future possibility of the needle device for the trace determinations of otherclasses of VOCs.

4. Conclusions
Several VOCs potentially related to a third-hand smoking hazard were successfully extracted
and determined with a polymer-packed extraction needle. The concentration monitoring of the time
variations for these smoking-related VOCs could allow for a more systematic evaluation of any
unpleasant odor and the suspected health hazard.
The present method [21] can also be applied to the determination of trace amounts of other
VOCs in environment air samples because of their simple and rapid extraction/desorption
procedure, along with an excellent repeatability and storage performance.

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