Environment International 98 (2017) 69–74

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Environment International

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Full length article

Higher dermal exposure of cashiers to BPA and its association with DNA
oxidative damage
Yanshan Lv a,1, Shaoyou Lu b,1, Yanyan Dai a, Caiyan Rui a, Yongjun Wang a, Yuanxiu Zhou a, Yanru Li a,
Qihua Pang a, Ruifang Fan a,c,⁎
a
Guangzhou Key Laboratory of Subtropical Biodiversity and Biomonitoring, School of Life Science, South China Normal University, Guangzhou 510631, China
b
Shenzhen Centers for Disease Control and Prevention, Shenzhen 518055, China
c
Guangzhou Key Laboratory of Environmental Exposure and Health, School of Environment, Jinan University, Guangzhou 510632, China

a r t i c l e i n f o a b s t r a c t

Article history: Bisphenol A (BPA) is a widely used chemical in the production of many polycarbonate plastics, epoxy resin linings
Received 16 July 2016 for food and beverage containers and thermal papers. Oral intakes from the contaminated diets were considered
Received in revised form 1 October 2016 as the predominant source of BPA exposure for humans. However, due to the high levels of BPA on thermal re-
Accepted 2 October 2016
ceipts and their wide applications in our daily life, the amount of BPA be transferred to the skin after holding ther-
Available online 8 October 2016
mal paper should not be underestimated, particularly for cashiers. To investigate the contribution of BPA
Keywords:
exposure levels via the dermal contact route and the relationship between BPA exposure level and oxidative
Bisphenol A DNA damage, six male volunteers were recruited and required to simulate the cashiers' work and handle the
Urine thermal receipts during the study period. Triclosan (TCS, an antimicrobial compound used widely in personal
8-hydroxy-2′-deoxyguanosine health and skin care products) was applied as a reference compound. Their urinary BPA, TCS and 8-hydroxy-
Dermal absorption 2′-deoxyguanosine (8-OHdG) concentrations were determined by high performance liquid chromatography/
tandem spectrometer (LC/MS/MS). The results showed that after handling the thermal receipts, the urinary
BPA concentrations of volunteers increased 3 times of those before the experimental period. But TCS levels in
urine kept stable. There existed a correlation between BPA exposure and 8-OHdG (R2 = 0.237, p b 0.001), but
not between TCS and 8-OHdG concentrations (R2 = 0.026, p b 0.777), indicating that more BPA exposure
could lead to higher oxidative DNA damage. That the increases in 8-OHdG levels in urine being almost consistent
with those of BPA suggested that handling thermal receipts resulted in the increasing BPA intakes and BPA expo-
sure was correlated with DNA oxidative damage. After 48 h of the end of handling thermal receipts, the urinary
BPA levels did not descend to the levels before experiment, suggesting that the excretion of BPA via dermal con-
tact was over 48 h. BPA exposure through dermal contact route contributed 51.9% to 84% to urinary BPA levels
with the GM ratio of 70.9% for cashiers, indicating that it might be seriously underestimated for cashiers accord-
ing to the previous studies. More attentions should be paid on the exposure of BPA via dermal penetration for
cashiers.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction 2013 (GrandViewResearch, 2014; Vandenberg et al., 2013). Contami-

Bisphenol A (BPA) is widely used in the production of many polycar-
bonate plastics, epoxy resin linings for food and beverage containers
and is confirmed with weak estrogenic activity in several in vitro and
in vivo preparations. Besides domestic plastic items, BPA is also used
as a preferred color developer in carbonless copy paper and thermal
paper. The production of BPA is reported to be 15 billion pounds in
⁎ Corresponding author at: Guangzhou Key Laboratory of Subtropical Biodiversity and
Biomonitoring, School of Life Science, South China Normal University, Guangzhou
510631, China.
E-mail address: 20001047@m.scnu.edu.cn (R. Fan).
1
Contribute equally to this work.
nated food and drinks by their packaging materials were believed to
be the major exposure route of BPA (Geens et al., 2012). However, the
concentrations of BPA in thermal paper were up to 20-30 μg/g paper,
and thousands times higher levels than those in foods and drinking
plastic packaging (Liao et al., 2012; Fan et al., 2015; Lu et al., 2013).
Due to its low cost, thermal papers containing BPA are still pro-
duced/used in massive quantities for a wide variety of commercial ap-
plications, including different cash receipts, luggage tags, faxes, and
labels. People frequently handle thermal papers in their daily life, partic-
ularly for cashiers. Biomonitoring data showed that BPA was detected in
more than 90% of all human urine samples (CDC, 2009; Pirard et al.,
2012; Völkel et al., 2011, Ndaw et al., 2016). Due to its weak estrogenic
activities, BPA exposure may influence multiple endocrine-related

http://dx.doi.org/10.1016/j.envint.2016.10.001
0160-4120/© 2016 Elsevier Ltd. All rights reserved.
70 Y. Lv et al. / Environment International 98 (2017) 69–74
pathways and is observed to be associated with prostate and breast can-
cer, neurobehavioral deficits, heart disease, and obesity (Bhandari et al.,
2015; McCaffrey et al., 2013). Furthermore, BPA may act as a DNA meth-
ylation agent and cause altered gene expression in the rodent brain
(Wolstenholme et al., 2011). Studies showed that BPA exposure is asso-
ciated with oxidation DNA damage (Watkins et al., 2015).
The BPA exposure routes via dermal contact and absorption could
not be ignored. BPA is readily absorbed and metabolized by the skin
and the dermal absorption ranged up to 46% efficiency (Zalko et al.,
2011). Physiologically based toxicokinetic modeling and simulated con-
centrations in blood, liver and kidney were used to assess the risk aris-
ing from the dermal exposure. The past results showed that the dermal
exposure can be assumed safe (Mielke et al., 2011). Demierre's studies
also conclude that dermal exposure to BPA contributes in a negligible
way to total body burden (Demierre et al., 2012). However, the statisti-
cal analysis of NHANES data suggested that individuals with potential
occupational exposure to thermal paper receipts are more likely to
have detectable levels of urinary BPA compared to individuals with
non-occupational exposure (p-value b0.001). Moreover, females with
the potential occupational exposure to thermal paper receipts may
have significantly higher levels of urinary BPA excretion compared to fe-
males with non-occupational exposure (Hehn, 2016). Even when using
a hand sanitizer contaminated by BPA after holding thermal receipt
paper could increase urinary and serum BPA levels immediately
(Hormann et al., 2014). Hence, the dermal penetration of BPA and its
contribution to BPA exposure are still issues to be resolved, particularly
for occupational exposed populations, such as cashiers. In conclusions,
BPA exposure through dermal contact might be underestimated and
the risks induced by non-oral BPA exposure routes should raise more
concerns.
To investigate the contribution of BPA exposure levels via dermal
contact route to human body burden and the relationship between
BPA exposure level and oxidative DNA damage, six male volunteers
were recruited and required to simulate the cashiers' work and handle
the thermal receipts during the study period. Triclosan (TCS, a phenol
compound whose chemical structure is similar to BPA and is a broad-
spectrum antimicrobial compound used in a variety of consumer prod-
ucts such as cosmetics) was used as a compound of reference in this
study. Their urinary BPA, TCS and 8-hydroxy-2′-deoxyguanosine (8-
OHdG, a sensitive marker of oxidative stress in urine) levels were deter-
mined by high performance liquid chromatography/ tandem spectrom-
eter (LC/MS/MS).
2. Materials and method
2.1. Chemicals and materials
BPA, TCS and D3-TCS were purchased from Dr. Ehrenstorfer (Augs-
burg, Germany). 8-OHdG was purchased from Sigma (St. Louis, USA).
13
C12-BPA and 15N5-8-hydroxy-2′-deoxyguanosine (15N5-8-OHdG)
were purchased from Cambridge Isotope Labs (MA, USA). The Helix
pomatia-derived β-glucuronidase/arylsulfatase was obtained from
Sigma (Santa Luis, USA). Methanol (LC-MS Chromasolv®, ≥99.9%) was
obtained from Fluka (USA). The water used for experiments was puri-
fied with a Milli Q system (Darmstadt, Germany). Glacial acetic acid, so-
dium acetate and KH2PO4 (HPLC grade) were obtained from Fisher
Scientific (Waltham, MA). All other reagents were analytical grade and
used without further purification. The Bond Elut C18 SPE cartridge
(500 mg and 6 mL) was obtained from Varian (Santa Clara CA, USA).
2.2. Subject recruitment and urine sampling
Six male volunteers (20–25 years, 170-175 cm in height, and 60–
65 kg in weight) from a university in Guangzhou, China, were recruited
to simulate the cashiers' work in July, 2014. The whole study spanned
over three periods, and lasted for 12 days in total. During the first five-
day period, the volunteers stayed in the university and did their own
routine work (not cashier's work) as usual. During the second five-day
period, the volunteers simulated the cashiers' work for 8 h each day.
They finished 30 times of check-out in 1 h, and touched the same ther-
mal paper used by cashiers with 23.5 mg BPA/g paper for 5 s each time.
The last two-day period, the simulation was completed, and they went
back to their own routine work as usual. Before eating meals/snacks,
they washed their hands with water for about 5 s as usual.
During the first five-day period (pre-experimental period), morning
urine samples were collected. During the simulation (experimental pe-
riod) and post simulation periods (post-experimental period), morning
(9:00 am), noon (13:00 pm) and night urine samples (17:00 pm) (4-h
intervals between the collections) were collected in screw-cap sealed
polyethylene bottles and the fresh urinary creatinine was determined
by spectrophotographic method (Taussky, 1954). One hundred and
fifty nine urine samples in total were collected. Urinary concentrations
of BPA, TCS and 8-OHdG were expressed as μg/L directly or corrected
by creatinine concentrations and expressed as μg/g crt.
Each participant was interviewed by a trained recruiter and was re-
quired to sign a consent form to take part in the study. Their personal in-
formation (i.e. age, height, weight, life habit and health condition) were
collected.
As diet, dust ingestion and dermal contact are the three exposure
routes to BPA, and in order to control the variations of BPA exposure
levels from dietary and dust ingestion during the experimental periods,
all the recruited students were provided the same foods and drinks by
the research group. Additionally, they were required to stay within
the lab areas, classrooms and dormitory in the university until the com-
pletion of the experiment.
2.3. The collection and measurements of BPA in dust
When BPA is released into environment, it is easily absorbed
through ingestion of dust. BPA exposure from dust ingestion is also an
important route of BPA intake (Wang et al., 2012). In order to control
and acknowledge the variations of BPA exposure from indoor and out-
door dust, BPA concentrations in dust were measured. Three indoor
spots (i.e. lab, classroom and dormitory) and four outdoor representing
spots which are located around the university campus were chosen.
Dust samples were collected in July, 2014.
Before extraction, dust samples were sieved and homogenized by a
2 mm sieve. Then 100 mg dusts were weighed and dissolved by 3 mL
of a mixture of methanol and acetonitrile (v:v = 1:1). After 10 min ul-
trasonic and 3 min vertex, the supernatants were taken out. The same
procedures were replicated for 3 times and all the supernatants were
combined. Then they were dried by gentle nitrogen gas and
reconstituted by 0.5 mL methanol and acetonitrile (v:v = 1:1). After fil-
tered by 0.22 μm filter membrane, they were stored under −20°Cuntil
analysis by high performance liquid chromatography coupled with fluo-
rescence detection (HPLC-FC). The instrument analysis was referred to
our previous report (Fan et al., 2015). The detection limit (LOD),
which were defined as 3 times signal-to-noise ratios (S/N) in methanol,
was 78 μg/L for BPA. The recoveries spiked in the QC samples were be-
tween 75 and 125% with RSDs of b 20%.
2.4. The sample preparation and measurements of BPA, TCS and 8-OHdG
The standard and sample preparation followed our previous method
(Fan et al., 2012). Briefly, a diluted urine pooled sample was used to pre-
pare the standard. The native standard concentrations were in the range
0.25–30 μg/L while internal standard concentrations were 50 ng/mL for
15
N5-8-OHdG, 10 ng/mL for 13C12-BPA and 25 ng/mL for D3-TCS. 300 μL
(high concentration, 15.0–37.5 μg/L), 100 μL (medium concentration,
5.0–12.5 μg/L) and 20 μL (low concentration, 1.0–2.5 μg/L) native stan-
dard mixtures were added into the same diluted pooled urine to pre-
pare the QA/QC samples. Standards (including blank and QA/QC
 Y. Lv et al. / Environment International 98 (2017) 69–74 71

samples) were prepared on the day of use. All the calibration standards,
blank and QA/QC samples were processed with the real samples in the
same batch run.
10 μL β-glucuronidase/arylsulfatase enzyme and 3 mL 0.1 mol/mL
HAC-NaAC buffers were added to 2.0 mL urine sample to adjust pH to
5.5, and then incubated at 37 °C overnight. The target analytes were ex-
tracted through solid phase extraction (SPE) cartridges. The SPE proce-
dure was followed our previous method (Fan et al., 2012).
BPA, TCS and 8-OHdG were simultaneously determined by a 20 A
HPLC (Shimadzu, Japan)/API Q-Trap 5500 mass spectrometer (AB,
SCIEX, USA). An XTerra MS C18 column (5 μm, 4.6 × 250 mm column,
waters, Massachusetts, USA) was employed to separate urinary metab-
olites. Each analyte was quantified by its own deuterated, 15N- or 13C-la-
beled internal standards. Electrospray ionization (ESI) was carried out
in negative mode. The source temperature was set at 550°C and the ion-
ization voltage was −4500 V. The analytes were quantified in multiple
reaction monitoring (MRM) mode with a dwell time of 50 ms. The neb-
ulizer gas (GS1) pressure and the turbo heater gas (GS2) pressure were
both 55 psi. The curtain gas (CUR) and the collision gas (CAD) pressure
was 20 psi and medium, respectively. The optimized parameters of
mass spectrometer were listed in Table 1.
0.1% acetic acid in water (v/v) and methanol were used as mobile
phases. Firstly, a gradient elution program was initiated with 5% meth-
anol and kept for 1.5 min. Then methanol ratio was increased from 5% to
10% in 1 min and continually increased to 50% in 1.5 min. Then metha-
nol ratio increased to 95% in 4 min and was kept for 4.5 min. The flow
rate was set at 0.6 mL/min and the column temperature was held at
40 °C. The MDLs, which were defined as 3 times S/N in the spiked
urine sample, were 0.20 μg/L for 8-OHdG, 0.63 μg/L for BPA, 0.31 μg/L
for TCS, respectively.
2.5. Calculation of BPA exposure from dust ingestion
Based on the median concentrations of BPA measured in dust sam-
ples, we estimated the daily intake (EDI; ng/kg bw/day) of BPA through
dust ingestion as shown in Eq. 1 (Liao et al., 2012):
C  DIR
EDI ¼
BW
Hence, we could calculate the contribution ratio of dermal contact to
urinary BPA and daily intakes of BPA for cashiers by Eqs. 2 and 3.
½BPAduring −½BPAbefore
Ratio ¼ Eq: 2
½BPAduring
EDI ¼ C  V=BW Eq: 3
Here, EDI represented the daily intake of BPA mainly from dermal
contact(μg/kg BW/day); C is the urinary BPA concentration (μg/L),V is
the urine volume for adult (assumed as 2 L for one day), BW represent-
ed body weight and assumed as 60 Kg for adults. We assumed if BPA en-
tered human body via dermal penetration 100% of them would circulate
in blood and would be excreted through urine.
2.7. Quality assurance and quality control (QA/QC) and data statistic
analysis
Method accuracy and precision were evaluated by replicate analysis
of matrix spikes at three levels of 15.0–37.5 μg/L, 5.0–12.5 μg/L and 1.0–
2.5 μg/L for urinary BPA, TCS and 8-OHdG. Recoveries were 80–120%
with relative standard deviations (RSDs) of 10–20%. Two methanol
blanks were run following the analysis of the highest levels of calibra-
tion standards and QC samples to examine and avoid the potential
carry-overs. In order to evaluate the precision and accuracy of the real
sample analysis, 15% of the real urine samples in each batch were ana-
lyzed in duplicates and the relative percentage difference (RPD) was
in a range of less than ±20%. After the analysis of every 10 urine sam-
ples, the lowest level of calibration standard was analyzed to check
the sensitivity of instruments.
SPSS (SPSS, version 20.0, Chicago, IL) was employed in the statistical
analysis of results. The skewness normality was applied to check the
distribution normality of target analyte concentrations. ANOVA test
was used to detect significant difference of geometric means (GMs)
among the volunteers during different periods. The level of significance
was set as p b 0.05. The Spearman correlation (one tailed) was taken to
test the associations between variables.
Eq: 1
3. Results

Where C is the concentration of BPA in dust samples (ng/g), DIR is
dust ingestion rate which is 200 (mg/day) for adults (U. S. EPA, 2012;
Jones-Otazo et al., 2005), and BW is body weight (assumed 60 kg for
an adult). We assumed an absorption efficiency of 100% for BPA from
dust to systemic blood circulation (Liao et al., 2012).
2.6. Calculation of contribution to urinary BPA from dermal contact and
daily intakes of BPA
There are three routes for human BPA exposure, i.e. diet, dust inges-
tion and dermal contact. Then if the exposure routes of diet and dust in-
gestion were controlled, then the BPA exposure for the volunteers
should be mainly from the dermal intakes of contacting thermal papers.
3.1. Levels of BPA in dust and EDIs of BPA through dust ingestion
Seven dust samples were collected, including three indoor and 4
outdoor dust samples. The results showed that the concentrations of
BPA in outdoor dusts were 1.05, 3.43, 1.54 and 3.49 μg/g with the GM
concentration of 2.38 μg/g and the levels of BPA in indoor dusts were
3.45, 1.21, 0.84 μg/g with the GM concentration of 1.83 μg/g. The EDI
via dust ingestion was 7.17 ng/kg BW/day for college students living
in the campus. Average contributions to urinary BPA via dust ingestion
before, during and after the experimental period were 3.77%, 2.39%
and 1.84%, respectively.
3.2. Concentrations of BPA, TCS and 8-OHdG at different experimental
periods

Table 1
The variations of urinary TCS concentration for volunteers before,
Optimized MS/MS parameters for each analyte. during and after the experimental period were limited (i.e. 1.96 ±
11.3 μg/g crt. & 5.94 ± 52.6 μg/L), (2.01 ± 10.8 μg/g crt. & 3.36 ±
Analytes RT/min Q1 Q3 DP (eV) EP (eV) CE (eV) CXP (eV)
21.9 μg/L and 2.03 ± 7.17 μg/g crt. & 2.95 ± 23.4 μg/L, respectively) and
8-OHdG 5.42 282 192 −110 −10 −27 −20 GM concentrations of TCS were almost kept stable for six recruiters. But
15
N5-8-OHdG 5.45 287 197 −96 −10 −28 −27
GM concentrations of BPA in urine varied greatly in different periods.
BPA 8.42 227 212 −100 −10 −25 −12
13
C12-BPA 8.44 239 224 −100 −10 −25 −13 Whether in μg/g crt. or in μg/L, significant differences were observed for
TCS 9.91 287 35 −100 −10 −40 −11 BPA levels and 8-OHdG concentrations among different periods
D3-TCS 9.93 290 35 −65 −10 −45 −10 (p b 0.05). Before the experiment, GM level of BPA exposure was
DP, declustering potential; EP, entrance potential; CE, collision energies; CXP, collision cell 1.89 ± 1.87 μg/g crt. for volunteers. Then GM BPA concentration went
exit potential. up to 8.20 ± 11.4 μg/g crt. during the experimental period, and was
72 Y. Lv et al. / Environment International 98 (2017) 69–74

Table 2
Concentrations of urinary BPA, TCS and 8-OHdG and EDI of BPA (μg/kg BW/day) for six volunteers during different experimental periods.

BPA EDIs of BPA TCS 8-OHdG

Compounds
GM ± SD Range GM ± SD Range GM ± SD Range GM ± SD Range

Pre-experimental period μg/g crt. 1.89 ± 1.87 0.02–7.42 0.18 ± 0.17 0.00–0.76 1.96 ± 11.3 0.02–40.3 6.35 ± 10.3 0.25–44.4
μg/L 5.44 ± 5.05 0.02–22.9 5.94 ± 52.6 0.06–189 18.3 ± 29.1 0.53–130
Experimental period μg/g crt. 8.20 ± 11.4 0.20–57.1 0.30 ± 0.87 0.01–4.93 2.01 ± 10. 8 0.04–58.3 31.7 ± 40.2 2.50–177
μg/L 29.4 ± 25.9 0.18–148 3.36 ± 21.9 0.06–132 51.8 ± 38.5 1.17–185
Post-experimental period μg/g crt. 8.10 ± 7.97 0.41–29.2 0.39 ± 0.51 0.01–1.75 2.03 ± 7.17 0.06–29.0 31.8 ± 33.7 5.19–144
μg/L 11.63 ± 15.2 0.19–52.4 2.95 ± 23.4 0.09–107 46.3 ± 38.5 5.64–149

almost 4 times compared to those before the experimental period. After were very similar to that of BPA concentrations without creatinine ad-
the experimental period, the urinary GM BPA concentration still stayed justment (μg/L, data not shown). Furthermore, the trends of 8-OHdG
in a high level with 8.10 ± 7.97 μg/g crt. Interestingly, concentrations of levels were almost the same as that of BPA, and the 8-OHdG levels
urinary 8-OHdG were 6.35 ± 10.3, 31.7 ± 40.2, 31.8 ± 33.7 μg/g crt., were elevated along with the BPA concentrations.
respectively with similar trends as BPA exposure levels (Shown in The Spearman correlation analysis showed that a correlation exists
Table 2). between urinary BPA exposure and urinary levels of 8-OHdG with a co-
efficient of 0.237 (p b 0.001), but no correlation was found between TCS
3.3. The association of BPA concentrations and 8-OHdG levels and 8-OHdG concentrations in urine (R2 = 0.026, p = 0.777).

The daily average concentrations of BPA, TCS and their correspond- 3.4. The contribution of dermal contact to urinary BPA exposure
ing 8-OHdG levels in urine for six volunteers were listed (Shown in
Fig. 1). In all the urine samples for all six volunteers, TCS concentrations The EDIs for BPA for the six volunteers were computed based on the
were kept almost stable except for one volunteer (i.e. person 2). Urinary urinary concentrations of BPA (Table 2). The EDIs of BPA for the differ-
BPA varied greatly between different experimental periods. Whether in ent experimental periods were 0.19 ± 0.22, 0.30 ± 0.87 and 0.39 ±
the unit of μg/g crt. or in μg/L, the urinary BPA concentrations always in- 0.51 μg/kg BW/day, respectively. Individual BPA exposure via thermal
creased after the experimental period. Though BPA levels for individuals contact accounted for 71.1, 51.9, 77.1, 81.7, and 64.7% of the urinary
were different, all six volunteers had a generally similar trend. BPA con- BPA with the GM ratio of 70.9 ± 12.2% (Table 3).
centrations were lower prior to the experimental period, and were al-
most kept at stable levels during most of the time, but they increased 4. Discussion
immediately after the first simulation day and kept at consistently
high levels till the end of the experimental period. Finally BPA concen- It is reported that dust ingestion, diet intake by BPA and dermal ab-
trations declined slightly or had high levels after the experimental peri- sorption are the three major exposure routes to BPA for human
od. The trends of urinary BPA concentrations (μg/g crt.) versus time (Myridakis et al., 2016). According to the measured BPA concentrations

Person 1 Person 2 Person 3

50 80
40
BPA
TCS BPA
BPA
8-OHdG TCS
TCS 40
30 60 8-OHdG
8-OHdG
Concentrations(µg/g crt)

Concentrations(µg/g crt)
Coccentrations(µg/g crt)

30

20 40

20

10 20
10

0 0
0

0 1 2 3 4 5 6 7 8 9 10 11 12 13 0 1 2 3 4 5 6 7 8 9 10 11 12 13 0 1 2 3 4 5 6 7 8 9 10 11 12 13

Days Days Days

Person 4 Person 5 Person 6

100
120
BPA 100
TCS BPA
BPA
80 8-OHdG 100 TCS
TCS
8-OHdG 80 8-OHdG
Concentrations(µg/g crt)

Concentrations(µg/g crt)

Concentrations(µg/g crt)

80
60
60
60
40
40
40

20
20
20

0 0
0

0 1 2 3 4 5 6 7 8 9 10 11 12 13 0 1 2 3 4 5 6 7 8 9 10 11 12 13 0 1 2 3 4 5 6 7 8 9 10 11 12 13

Days Days Days

Fig. 1. Daily distributions of urinary BPA, TCS and 8-OHdG concentrations of the six volunteers (unit: μg/g crt.)
 Y. Lv et al. / Environment International 98 (2017) 69–74
Table 3
The contribution ratios of dermal contact to urinary BPA exposure for six volunteers.
Person 1 2 3 4 5 6
EDI before simulation 3.44 2.98 2.37 2.75 1.53 4.10
EDI during simulation 11.9 6.19 10.4 18.0 8.38 11.6
Ratio (%) 71.1 51.9 77.1 84.8 81.7 64.7
GM ± SD of Ratio (%) 70.9 ± 12.2
in dust from the campus, they were in the medium levels compared to
previous reports (Liao et al., 2012; Geens et al., 2009). The calculated
contribution ratios of dust ingestion to the urinary BPA based on EDI
data were in the range of 1.84–3.77%. Hence, the dust ingestion expo-
sure route to BPA for college students could be ignored in our simulation
experimental study, dermal absorption and diet intakes were the main
routes of BPA exposure for the volunteers. Based on that the diets in-
cluding drinking and food were strictly controlled during this study,
though we did not quantify the daily dietary intakes of BPA, the varia-
tions of BPA intakes from diets during study period were well con-
trolled. Therefore, any differences and/or variations in urinary BPA
levels observed in the volunteers might be from the incurrence of han-
dling thermal receipts with high levels of BPA.
TCS is a widely used antimicrobial compound in cosmetics and per-
sonal health care products, such as lotion, shampoo, toothpaste et al.
and dermal absorption is its main exposure route for human. As it has
wide applications in human daily lives, it is often detected in human
urine (Mortensen et al., 2014). Furthermore, because its chemical struc-
ture is similar to BPA and no TCS exits in thermal receipts, it was used as
a reference compound in this experiment to indicate the dermal intakes
of BPA. Except for one person, urinary TCS concentrations in all the other
volunteers were stable during the whole study and no difference in var-
iations was found before, during and after the experimental periods.
After a comprehensive investigation of his life habits during the study,
we were informed that the volunteer used a large quantity of skin
care products and changed skin and hair care products during the
study. This change of products could be a reasonable explanation as to
why his average urinary TCS concentration (20.6 μg/g crt.) was signifi-
cantly higher than those of the other volunteers (3.22 μg/g crt.).
For the urinary BPA levels, the GM concentration during the experi-
mental period was significantly higher than that before the experimen-
tal period, indicating that the increasing BPA amounts were from
handling thermal receipts. The varied EDIs between different periods
also suggested that dermal contact was the main route of BPA intake
for cashiers. As thermal receipts are used in many supermarkets and
stores due to their low price, cashiers might absorb considerable BPA
though skin every day. Moreover, because most cashiers in China are
young girls and boys who are in a stage of marriage (forming a family)
and pregnancy (productive period), the potential health risks from BPA
exposure could not be underestimated or ignored.
It is well known that the excretion of BPA via urine is quick and
the metabolite time does not exceed 24 h (Geens et al., 2012;
Taylor et al., 2011). Interestingly, though the handling of thermal re-
ceipts ended in 48 h, the urinary BPA levels did not decrease to the
levels before the experimental period. On the contrary, some even
increased slightly. We speculated that the difference of bio-availabil-
ity between oral and dermal route, as well as BPA trapped by the
palm skin might lead to the lag time of BPA intake via dermal contact.
Firstly, the path and metabolism of oral intakes and dermal absorp-
tion were different. Parts from oral intake could easily enter the
blood stream and be excreted by the liver. But parts of dermal ab-
sorption intake should overcome the penetration of skin and then
enter the blood stream. Additionally, the surface of palm skin has
many folds and wrinkles which could trap a large fraction of BPA
even after handwashing. Though washing hands could reduce BPA
dermal exposure, there is still about 19–47% amount of BPA left on
the hands after washing (Fan et al., 2015).
73
Many studies of BPA biomonitoring focus on sensitive populations,
such as infants, children and pregnant women due to its estrogenic ef-
fects (Myridakis et al., 2016; Mortensen et al., 2014). But little attention
has been paid to occupationally exposed populations like cashiers. The
results of this showed that the concentrations of urinary BPA during
handling thermal receipts covered three magnitudes, and were 3–20
times higher than those before simulation. This might be because BPA
intake via dermal contact was strongly with the touching strength,
touching areas of the palms and touching time (Fan et al., 2015). Due
to the significant increase of BPA exposure during work time for ca-
shiers, we suggested that cashiers should wear gloves and wash hands
carefully with lotions or soaps after work to avoid of more BPA intakes
from the dermal contacts.
Porras et al. studied BPA exposure via thermal paper receipts in sim-
ulation experiments performed by three volunteers, and examined the
urinary excretion of BPA. They concluded that it was safe for cashiers be-
cause the total daily intake over was 25 times lower than the European
Food Safety Authority's (EFSA's) proposal for a temporary tolerable
daily intake (TDI) (Porras et al., 2014). Our results showed that with in-
creasing BPA levels, urinary 8-OHdG increased, suggesting that a higher
BPA intake could lead to higher oxidative DNA damage. The Spearman
correlation analysis also confirmed that 8-OHdG levels correlated with
BPA exposure levels in urine, but TCS did not. All these indicated that
BPA exposure may lead to potential health risks, particularly for those
occupational populations that have to deal with thermal paper contain-
ing BPA. Although the EDIs in the study were lower than the reference
values given by the U.S. Environmental Protection Agency (U.S. EPA,
50 μg/kg BW/day) or the EFSA's proposal for a temporary TDI (5 μg/
kg/day) (U. S. EPA, 2008; ESFA, 2006), as the BPA intakes were low
and chronic for cashiers, the potential health risk still could not be ig-
nored. Mounting evidence has suggested that the low dose of BPA expo-
sure plays an important role in the prevalence of existing metabolic
syndromes (Rubin et al., 2001; Wei et al., 2011).
Human skin in an in vitro test showed that 8.6% of BPA via dermal
contact could penetrate the skin and the amount of bio-available BPA
after 24 h exposure reached 0.169 g/cm2, accounting for 9.3% of the
total dose applied. Therefore, it confirmed that systematic exposure to
BPA via skin contact contributed in a negligible way to the total BPA ex-
posure (Demierre et al., 2012). Zalko examined the diffusion and me-
tabolism of BPA using viable skin models and observed that BPA is
readily absorbed and metabolized by the skin and the absorption ratio
is 27% in humans (Zalko et al., 2011). Nevertheless, the average contri-
bution ratio of BPA via dermal contact to urinary BPA in our study
showed that dermal absorption was not a route which could be ignored
for BPA exposure, accounting for 70.9% contribution ratio to urinary
BPA. The health risks might be well underestimated for cashiers' previ-
ous studies. Our results were in agreement with Gundert-Remy et al.,
who observed that if the absorption on the oral route is b 100% the pro-
cedure may underestimate the risk of the exposures of the non-oral
route (Gundert-Remy et al., 2013).
5. Conclusions
BPA could be absorbed via dermal contact, and handling thermal re-
ceipts is an important route of BPA exposure for cashiers. Urinary BPA
levels increased significantly due to the dermal contact of receipts
made with high BPA concentrations. The intake and metabolic time of
the dermal absorption of BPA lagged over 48 h compared to the oral in-
take. A higher BPA intake resulted in higher oxidative DNA in urine. The
contributions of non-oral routes of BPA intake ranged from 51.9% to
84.8% for cashiers and the contribution ratio of dermal contact to uri-
nary BPA was seriously underestimated. Although the EDIs of the volun-
teers were considered safe and lower than the reference values given by
EPA or ESFA, more attentions should be paid to cashiers because their
BPA exposure is long term and accumulated.
74 Y. Lv et al. / Environment International 98 (2017) 69–74
Acknowledgements
We thank for the financial supports from Science and Technology
Program of Guangzhou, China (Grant No. 201510010107), Guangzhou
Key Laboratory of Environmental Exposure and Health (Grant No.
GZKLEEH201607) and Guangdong Natural Science Foundation (No.
S2013010013183).
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