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Occurrence of bisphenol A in surface water, drinking water and plasma from Malaysia
with exposure assessment from consumption of drinking water
V.A. Santhi
a
, N. Sakai
a,b
, E.D. Ahmad
a
, A.M. Mustafa
a,
⁎
a
Shimadzu_UMMC Centre of Xenobiotic Studies, Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia
b
Department of Environmental Engineering, Graduate School of Engineering, Kyoto University, 6158540 Kyoto, Japan
abstractarticle info
Article history:
Received 25 January 2012
Received in revised form 16 April 2012
Accepted 16 April 2012
Available online 9 May 2012
Keywords:
Bisphenol A
River water
Drinking water
Plasma
Exposure assessment
This study investigated the levelof bisphenol A (BPA) in surface water used as potablewater, drinking water(tap
and bottled mineral water) and human plasma in the LangatRiver basin, Malaysia.BPA was present in 93% of the
surface water samples at levels ranging from below limit of quantification (LOQ; 1.3 ng/L) to 215 ng/L while six
fold higher levels were detectedin samples collected near industrial and municipal sewage treatment plant out-
lets. Low levels of BPA were detected in most of the drinking water samples. BPA in tap water ranged from 3.5 to
59.8 ng/L with the highest levels detected in samples collected from taps connected to PVC pipes and water filter
devices. Bottled mineral water had lower levels of BPA (3.3± 2.6 ng/L) although samples stored in poor storage
condition had significantly higher levels (11.3± 5.3 ng/L). Meanwhile, only 17% of the plasma samples had de-
tectable levels of BPA ranging from 0.81 to 3.65 ng/mL. The study shows that BPA is a ubiquitous contaminant
in surface, tap and bottled mineral water. However, exposure to BPA from drinking water is very low and is
less than 0.01% of the tolerable daily intake (TDI).
© 2012 Elsevier B.V. All rights reserved.
1. Introduction
Bisphenol A (BPA; 4, 4′-dihydroxy-2, 2-diphenylpropane; CAS# 80-
05-7) is a monomer used extensively in the production of polycarbonate,
epoxy resins and as a non-polymer additive in plastics such as polyvinyl
chloride (PVC), and water pipes (Welshons et al., 2006). Thus, the global
demand for BPA is expected to grow from 3.9 million tons in 2006 to
about 5 million tons in 2010 (Tsai, 2006). It is frequently detected in
wastewater discharged from municipal and industrial sources at levels
ranging from 0.23 to 149 μg/L (Fuerhacker, 2003; Lee et al., 2002;
Suzuki et al., 2004; Höhne and Püttmann, 2008; Sanchez-Avila et al.,
2009). Studies conducted in Greece and Spain have shown that only
about 68 to 87% of BPA is removed by the applied sewage treatment pro-
cesses (Stasinakis et al., 2008; Sanchez-Avila et al., 2009) while the rest
ends in the receiving surface and coastal water (Basheer et al., 2004;
Chen et al., 2010).
Although BPA is rapidly biotransformed and excreted in the
urine, levels of unconjugated BPA detected in biomonitoring studies
suggest the human populations are at risk from internal exposure
(Vandenberg et al., 2010). BPA has a much lower affinity for nuclear es-
trogen receptors compared to 17β-estradiol. However, it causes similar
changes in some cell functions at levels between 1 pM and 1 nM (0.23
to 230 pg/mL culture medium) which are below the mean and median
levels of unconjugated BPA measured in blood and tissues (Welshons
et al., 2006). Furthermore, Hugo et al. (2008) found BPA at 0.1 and
1 nM more effective at inhibiting the release of a key adipokine that pro-
tects humans from metabolic syndrome. Higher exposure to BPA may
also be associated with chronic health conditions such as diabetes and
cardiovascular disease in humans (Lang et al., 2008).
Exposure to this endocrine disruptor in humans is mainly through
food (Vandenberg et al., 2007) although Stahlhut et al. (2009) suggests
other nonfood exposures such as residential water supply. Stackelberg
et al. (2004) had previously reported BPA in drinking water at levels
as high as 0.42 μg/L while consumption of bottled mineral water may
also contribute to the exposure of endocrine disruptors (Wagner and
Oehlmann, 2009). The current tolerable daily intake (TDI) established
by the European Food Safety Agency (EFSA) for BPA at 50 μg/kg (body
weight)/day is similar to the reference dose (RfD) established by the
US Environmental Protection Agency (EPA). However, many studies
have shown that the TDI value is orders of magnitude greater than the
levels found to produce adverse endocrine effects (Ginsberg and Rice,
2009; Kang et al., 2006).
The study on BPA in water, especially when it's used for human con-
sumption is important. The rivers in Malaysia supplies over 98% of the
nation's potable water supply which are mostly treated using conven-
tional water treatment processes. However, many of these rivers are re-
cipients of treated effluents, municipal and industrial waste (Fulazzaky
et al., 2010). Additionally, there are reports of incomplete removal of
many pharmaceuticals and organic wastewater contaminants including
BPA during these treatment processes (Kim et al., 2007; Stackelberg et
al., 2007; Huerta-Fontela et al., 2008). In the present study, we analyzed
BPA level in river water which is used as potable water supply. We also
Science of the Total Environment 427–428 (2012) 332–338
⁎Corresponding author. Tel.: + 60 3 79492103; fax: +60 3 79674791.
E-mail address: mustafa@ummc.edu.my (A.M. Mustafa).
0048-9697/$ –see front matter © 2012 Elsevier B.V. All rights reserved.
doi:10.1016/j.scitotenv.2012.04.041
Contents lists available at SciVerse ScienceDirect
Science of the Total Environment
journal homepage: www.elsevier.com/locate/scitotenv
Author's personal copy
collected samples from outlets of municipal sewage treatment plants,
industryand wet market to assess the pollution load from these sources.
Furthermore, BPA levels in tap water, and bottled water and the influ-
ence of poor storage condition were studied with the aim of assessing
the potential human exposure from these sources. Finally, we measured
the internal exposure to unconjugated BPA in adults from selected
communities.
2. Materials and methods
2.1. Sample collection
2.1.1. River water
The Langat River basin covers an area of approximately 2400 km
2
.
Its three major tributaries, Langat, Semenyih and Labu River flow
through the states of Selangor, Negeri Sembilan and Federal Territory
of Putrajaya and Cyberjaya. These rivers provide potable water supply
to the people living in the basin and nearby Putrajaya, Cyberjaya and
Kuala Lumpur, the capital. During the selection of samplingsites, special
emphasis was given to the intake points of drinking water treatment
plant (DWTP) located in the basin. These samples will represent the
quality of raw water abstracted by the respective treatment plants.
Samples were collected with the assistance of district health staff that
has access to the restricted intake point area. Accordingly, monthly
samples were collected from the seven sites from September 2008 to
July 2009 (Fig. 1) except for the months of October 2008 and March
2009 when the health staff were not available. In addition, only 4 sam-
ples were collected from site 3 due to the shutdown of the DWTP from
high turbidity, thus preventing access to the sampling site. One sample
was not collected from site 6 due to heavy rain on the sampling day.
Two liters of samples was collected in thoroughly cleaned amber
glass bottles. These bottles were rinsed with sample water before filling
to the brim and capping with an aluminum foil lined cap. In situ mea-
surements of turbidity using Hach 2100P and pH with temperature
NEGERI
SEMBILAN
12
3
4
5
6
7
Straits of Malacca
Batang Labu River
SELANGOR
NEGERI
SEMBILAN
Nilai Industrial
Park
2
3
Bandar Baru
Nilai
Intake of
Salak Tinggi
WTP
Agricultural area
1
4
5
Fig. 1. Location of sampling sites in Langat River basin. Sampling for point source pollution source was carried out upstream of Batang Labu River, along Batang Nilai River.
333V.A. Santhi et al. / Science of the Total Environment 427–428 (2012) 332–338
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using Hach One were recorded. Samples were preserved from microbial
activity by acidification with nitric acid and transported to the laborato-
ry in a cool box at 4 °C. They were then filtered with Whatman GF/B
(1.0 μm) and GF/F (0.7 μm) glass fiber filters and stored in the dark at
4 °C. All samples were analyzed within 48 h of collection.
Average monthly rainfall data from Kg. Salak rainfall station was
provided by the Department of Irrigation and Drainage (DID). This
data was used to assess the seasonal trend of BPA occurrence in the
basin. The months of September and November in 2008 and the months
of April and May in 2009 which receive monthly rainfall above 100 mm
were categorized as rainy months. As the average rainfall in December
2008 was only 25 mm compared to the usual 100 to 200 mm, it was cat-
egorized as a dry month as with the other months.
2.1.2. Point source pollution sources
Sampling for point source pollution sources was conducted up-
stream of Batang Labu River, along Batang Nilai River in December
2009 (Fig. 1). Domestic and industrial wastewater from Nilai town
and Nilai Industrial Estate, respectively flow into the Batang Nilai
River and subsequently into Batang Labu River. The distance from
Nilai town to the intake point of Salak Tinggi DWTP is about 13 km.
This WTP has experienced numerous shut downs due to high levels of
ammonia, phenolic compounds and smell (PNSB, 2009, 2010). In 2009
alone, Salak Tinggi DWTP was shut down for a total of 2480 h due to
poor water quality (PNSB, 2010). Single samples from outlets of munic-
ipal sewage treatment plants (sites 2, 3 and 5), Nilai industrial estate
(site 1) and wet market (site 4) were collected. Dissolved oxygen
level using Hach Oximeter was also measured together with the other
in situ parameters.
2.1.3. Tap water
Thirty tap water samples (samples coded 1 to 30) were collected
from houses in Kuala Lumpur and the surrounding area which share
the same source water from Langat River. The samples were collected
from the kitchen tap which receives direct water supply from the
main distribution pipe. Chlorine level was recorded using a Hach pocket
colorimeter before adding ascorbic acid to quench it. All the samples
were analyzed within 24 h.
2.1.4. Bottled mineral water
Nine brands of mineral water bottled in PET containers were pur-
chased from shops around Kuala Lumpur. The source water is filtered
ground water abstracted locally and is either ozonated or ultraviolet
(UV) irradiated after bottling. The water was packaged in 500 mL bot-
tles and four bottles of each brand were obtained. The single samples
were either analyzed upon purchase or after storage in an oven at
50 °C for 3 days. This is to simulate the mode of storage by some distrib-
utors in non-air conditioned stores where temperatures can reach up to
45 °C.
2.1.5. Blood plasma
For an initial screening of plasma BPA level, 101 random samples
were collected from communities living in the Langat River basin
which are expected to consume water from the same source. The age
of study subjects ranged from 19 to 69 years. Of these, 45.5% were
males and 54.5% females with an average age of 45±12 and 41±
11 years, respectively. Prior approval was obtained from the Ethics
Committee of University of Malaya Medical Centre (UMMC) for the col-
lection and analysis of human specimen. The samples were collected in
heparinized tubes using glass syringe. They were then centrifuged at
3500 rpm for 10 min and the supernatant stored in glass vials at
−20 °C before extraction.
2.2. Standards and reagents
The solvents (methanol, acetone, hexane, dichloromethane and
ethyl acetate) were of GC grade and purchased from Merck (Darmstadt,
Germany). They were further distilled in an all glass apparatus prior to
use. Analytical standard BPA and internal standard BPA-d
16
were sup-
plied by Supelco (USA). Stock solutions of 1000 mg/L of both standards
were prepared in methanol and stored at −20 °C. Working standardsat
various levels for spiking and preparation of calibration curve were pre-
pared daily. Silylating agent N,O-bis (trimethylsilyl) trifluoroacetamide
(BSTFA) with 1% trimethylchlorosilane (TMS) was supplied by Waco
(Japan). Nitric acid was supplied by Merck (Darmstadt, Germany)
while ascorbic acid by John Kollin Chemicals (UK). Sodium dihydrogen
orthophosphate used to prepare phosphate buffer was obtained from
Ajax Finechem (NSW, Australia) while phosphoric acid was from Fisher
Scientific (Leichestershire, UK).
Silica based bonded C18 (EC) cartridges (1 g/6 mL for water samples
and 500 mg/6 mL for plasma samples) were obtained from Biotage,
(EU). “Sole”brand mineral water from Italy was used for method vali-
dation and preparation of calibration curve for water samples. Expired
blood plasma obtained from the UMMC blood bank was treated with
activated carbon and used as blank sample.
2.3. Extraction method and analysis
2.3.1. Extraction and analysis of water sample
Prior to extraction, the pH of 1 L water sample was adjusted to 2–3
using nitric acid (1:1, v/v), spiked with 50 ng internal standard, BPA-d
16
before passing through the C18 cartridge. The cartridge was first rinsed
by passing 6 mL of acetone:hexane (1:1), followed by 10 mL of methanol
and 10 mL of water at a flowrateof1mL/min.Thesamplewasthenper-
colated through the cartridge at a rate of 4 mL/min, rinsed with 6 mL of
water and dried under high vacuum for 30 min. Elution was performed
with 4 ×2.5 mL of acetone:hexane (1:1). The eluant was dried under a
gentle stream of nitrogen, reconstituted with 100 μLofacetone:hexane
(1:1) and 20 μL of BSTFA +1% TMS before derivatizing at 75 °C for
40 min. One microliter was injected to the GC/MS for BPA analysis.
A Shimadzu QP-2010 GC/MS coupled with an auto sampler
(Shimadzu AOC-20S) was employed for the analysis. The target com-
pound was separated using a SGE BP-1 (Australia) capillary column
(length: 30 m; i.d.: 0.25 mm; film thickness 0.25 μm). Conditions for
GC/MS were as previously reported by Santhi et al. (2012).
2.3.2. Extraction and analysis of plasma sample
The target compound was extracted from 1 mL of plasma using a
method previously reported by Tan and Ali Mohd (2003). Briefly,
1 mL of sample was spiked with 5 ng of BPA-d
16
before loading on
the conditioned cartridge. The cartridge was then washed with 2 mL
of buffer and dried under vacuum for 10 min. BPA was eluted with
3 mL of dichloromethane:ethyl acetate (1:1) and dried under a gentle
stream of nitrogen. Prior to analysis, it was derivatized with 20 μLof
BSTFA+ 1% TMS at 75 °C for 40 min, dried under nitrogen and finally
reconstituted in 100 μL dichloromethane:ethyl acetate (1:1). One
microliter was injected to the GC/MS for analysis.
2.4. Statistical analysis
Statistical analysis was conducted using Statistical Package for the
Social Sciences (SPSS) for Windows, Version 15.0 (SPSS Inc., Chicago,
IL, USA). Analytical results below the quantitation limit (LOQ) were uti-
lized while results below detection limit (LOD) were set equal to zero.
Differential statistics was used to calculate mean, median, standard de-
viation and range. BPA levels during the rainyand dry months, different
storage temperatures, in male and female plasma and tap water from
taps with and without additional fittings were compared using t-test.
Statistical significance was set at pb0.05.
334 V.A. Santhi et al. / Science of the Total Environment 427–428 (2012) 332–338
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2.5. Quality assurance and quality control
Since BPA is a ubiquitous contaminant in the laboratory environ-
ment, special care was takento avoid contact with plastic materials dur-
ing sample collection, extraction and analysis. For each batch of water
and plasma analysis, a procedural blank and spiked blank at different
concentrations were included. The BPA level in the procedural blanks
was below the detection limit. Recovery of internal standard which
also acts as surrogate was checked.
2.6. Method validation
The calibration curve was linear over the selected range with corre-
lation coefficient (R
2
)>0.995 for both the water and plasma samples.
Water spiked with BPA at three levels (10, 100 and 500 ng/L) in repli-
cates of five was employed for the recovery, precision and accuracy
test. The average recovery forBPA in the range of 82 to 86% was satisfac-
tory. The precision of the method represented by percent relative stan-
dard deviation (RSD) was good and ranged from 3.1 to 8.7% (intra day)
and from 2.6 to 7.6% (inter day) while accuracy, expressed as percent-
age of bias ranged from −4.2 to 1.8%. Meanwhile, the recoveries for
plasma at 0.75, 8 and 20 ng/mL ranged from 80% to 97%. The intra day
and inter day precision ranged from 2.9 to 7.0% and from 2.7 to 8.9%,re-
spectively while bias ranged from −0.4 to 8.8%. The LOD was set as the
lowest level with a signal-to-noise ratio (S/N) of at least 3, while the
LOQ was 10 times S/N. The LOD values were 0.4 ng/L and 0.25 ng/mL
while the LOQ values were 1.3 ng/L and 0.75 ng/mL for water and plas-
ma samples, respectively.
3. Results and discussion
3.1. BPA in source (river) water
Levels of BPA detected in the source water samples are shown in
Table 1. BPA was detected in 93% of all the samples although detection
frequencies at upstream sites 1, 2 and 3 were lower. Levels of BPA varied
a lot among the sites ranging from below quantitation limit to 215 ng/L.
However, repeated sampling at the seven sites showed a clear difference
in the levels detected at each site. Higher levels were detected at sites 4,
5, and 6 which are located along the main Langat River and site 7 at its
tributary, Labu River. Although there are no industries upstream of
site 4, direct discharges into the river from houses and villages located
nearby were observed during a survey of the study area. Meanwhile,
the source water from sites 5, 6 and 7 receives effluents from numerous
sewage treatment plants, small and medium industries, housing estates
and towns located upstream. Site 6 which has the highest level of BPA
is located furthest down and thus receives effluents not only from up-
stream but also from the surrounding area. Site 7 while located in the vi-
cinity of palm oil plantations, is also located downstream of industries,
housing estates and townships.
There is no significant difference in the mean BPA level detected
during the rainy and dry months. However, when mean BPA for the
more polluted sites (5, 6 and 7) were compared, a significantly higher
level (p= 0.015) was detected during the dry months suggesting occur-
rence of dilution during the rainy months. The low levels of BPA
detected at upstream sites 1, 2 and 3 were comparable to the levels
(7.4 to 10.8 ng/L) measured upstream of Salut River in Tuaran, Malaysia
(Duong et al., 2010) while levels from urban sites 4 to 7 were compara-
ble to the levels (9 to 272 ng/L) reported by references in other surface
water (Bolz et al., 2001; Jin et al., 2004; Voutsa et al., 2006).
3.2. BPA in point source pollution sources
Results for BPA point source pollution sources are shown in Table 2.
The sample collected near the Nilai Industrial Estate (site 1) outlet had
1218 ng/L of BPA while the three samples collected near outlets of sew-
age treatment plants had a mean of 954± 299 ng/L. The BPA level in the
sample collected downstream of the wet market was also higher than
the levels detected in source water. Fifty‐four percent of the 17,600
water pollution point sources identified by the Department of Environ-
ment (DOE) in the country were sewage treatment plants while
manufacturing and agro based industries contributed another 39% and
3%, respectively (DOE, 2009). The discharged effluents from less than
75% of these sewage treatment plants comply with the Environmental
Quality (Sewage and Industrial Effluents) Regulation, 1979 (DOE,
2010). Additionally, 98% of the 1153 approved contravention licenses
granted under the Environmental Quality Act, 1974 for discharges into
water courses were given to sewage treatment plants (DOE, 2010).
Table 1
Levels of BPA detected at each sampling site during the sampling period (September to December 2008 and Jan to July 2009). Results are reported in ng/L.
Sampling month Sampling site
Site 1 Site 2 Site 3 Site 4 Site 5 Site 6 Site 7
September bLOQ
a
ND ND 16.1 8.9 4.0 14.3
October –––– – – –
November 2.5 13.8 –27.9 8.8 65.9 57.5
December 2.0 bLOQ –9.0 bLOQ 25.6 35.0
January 7.6 ND –6.4 21.8 16.3 171.0
February bLOQ ND –19.0 13.9 98.9 123.3
March –––– – – –
April ND ND –35.6 52.4 159.9 27.0
May 1.9 3.8 2.1 14.2 91.3 214.5 22.8
June ND 1.4 bLOQ 13.2 89.4 115.2 101.4
July 4.9 1.8 2.3 13.9 56.7 –203.0
Mean±SD 2.3±2.49 2.4 ± 4.45 1.4 ± 1.07 17.3 ± 9.20 38.2 ± 35.28 87.5 ± 74.27 83.9 ± 69.40
Median 1.9 bLOQ 1.6 14.2 21.8 82.4 57.5
Range ND–7.6 ND–13.8 ND–2.3 6.4–35.6 bLOQ–91.3 4.0–214.5 14.3–203.0
ND = not detected.
a
Below limit of quantitation (LOQ= 1.3 ng/L).
Table 2
Basic water parameters and BPA levels for point source pollution sources.
Site pH Turbidity
(NTU)
a
Dissolved oxygen
(mg/L)
Temperature
(°C)
BPA
(ng/L)
1 7.05 26.2 5.5 27.1 1218
2 7.04 20.5 3.5 28.1 1278
3 6.92 17.3 4.8 28.3 1015
4 7.10 25.8 5.4 28.3 581
5 7.31 22.1 5.3 29.1 629
a
NTU = nephelometric turbidity unit.
335V.A. Santhi et al. / Science of the Total Environment 427–428 (2012) 332–338
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Meanwhile, the main polluting industries in Langat basin were identi-
fied as plastic and PVC, engineering products, wood and paper products,
textile and electrical items (Osman et al., 2012). BPA is used as a plasti-
cizer in many of these products. The biological oxygen demand (BOD)
load from partially treated sewage and discharges from agro-based
and manufacturing industries in Selangor was 370,000 kg/day (DOE,
2009). These would suggest that industrial, domestic and sewage efflu-
ents are important anthropogenic sources of BPA in the basin.
3.3. BPA in tap water
Most of the DWTPs in Malaysia use conventional water treatment
processes comprising aeration, coagulation and flocculation, sedimen-
tation, sand filtration and chlorination. BPA was detected in all the tap
water samples at levels ranging from 3.5 to 59.8 ng/L (mean: 14.1±
14.36 ng/L) with twenty‐four samples having residual chlorine levels
ranging from 0.05 to 1.57 mg/L (Fig. 2). The finished water is usually
chlorinated at levels between 2 and 3 mg/L to ensure free residual chlo-
rine of not less than 0.2 mg/L throughout the distribution system (MOH,
2000). Although BPA rapidly degrades in highly chlorinated water, no
significant correlation was found when a Pearson correlation test was
performed between the levels of BPA and residual chlorine.
Samples 6 and 24 collected from taps fitted with filter devices and
sample 12 collected from a tap connected with PVC pipe had the highest
BPA levels at 56.4, 59.8 and 47.1 ng/L, respectively (mean; 54.4±
6.57 ng/L). Meanwhile, mean BPA level from the 27 samples collected
from taps without filter or PVC fittings was 9.6± 4.25 ng/L. Significant
difference was detected between these mean levels (pb0.01)
suggesting contamination of supplied water from these devices. While
water passingthrough PVC hose was previously reported to be contam-
inated by BPA (Yamamoto and Yasuhara, 2000), contamination from fil-
ter devices deserves further investigation as its usage is popular
especially in the urban areas of Malaysia (Aini et al., 2007).
The DWTP represented by site 4 is the largest treatment plant in the
study area with a capacity to produce 386 million liters per day (MLD)
of treated water. Most of the houses where the samples were collected
from were supplied by this DWTP. Mean BPA in tap water without filter
or PVC fittings was lower than source water for this DWTP (17.3±
9.20 ng/L) suggesting that conventional water treatment process was
able to remove some of the BPA. Similarly, in a study by Stackelberg et
al. (2007), 76% of BPA was removed from source water by conventional
treatment processes. The BPA levels in tap water from this study were
higher than the levels detected in Europe ranging from not detected
to 2 ng/L (Kuch and Ballschmiter, 2001; Loos et al., 2007) although it
was lower than the average 160 ng/L detected in Brazil (Sodre et al.,
2010) or the median 99 ng/L detected during summer time from
Guangzhou, China (Li et al., 2010).
3.4. BPA in bottled mineral water
Levels of BPA in mineral water bought off the shelf (storage temper-
ature, 25 °C) and after storage at 50 °C for 3 days to simulate improper
storage are shown in Fig. 3. Samples stored at room temperature had
lower levels of BPA (3.3± 2.6 ng/L) compared to those stored at higher
temperature (11.3± 5.3 ng/L) with significant difference detected be-
tween the mean levels. BPA was previously detected in all bottled min-
eral water samples from Japan although there were no increase in its
level after storage at 50 °C for 8 h (Toyo'oka and Oshige, 2000). Mean-
while, Casajuana and Lacorte (2003) detected a slight increase in
water bottled in PET containers after storage for 10 weeks. Mineral
water is described as ground water obtained from subterranean strata
through a spring, well, bore or other exit under The Malaysian Food
Regulations, 1985. Thus, the presence of BPA is most likely from con-
tamination during the production process involving extraction and fil-
tration of ground water. However, the increase in BPA after storage at
high temperature suggests contamination from the packaging material
itself. While plasticizers such as BPA and phthalates are not necessary
for the manufacturing of PET bottles, Bach et al. (2012) suggests that
usage of recycled PET may contributeto BPA contamination. In addition,
the mineralbottle closures may be a source of BPA in this study assam-
ples were stored on their side in the oven, with water having direct con-
tact with theplastic closures. BPA level in mineral water from this study
is similar to the level reported in bottled water from Greece (median:
4.6 ng/L) (Amiridou and Voutsa, 2011) although Shao et al. (2005)
found no detectable BPA in mineral water from China.
3.5. BPA in plasma
BPA was detected in 17% of the plasma samples at levels ranging
from 0.81 to 3.65 ng/mL. A similar low frequency of detection was
found in the serum of a Chinese population (n= 952) without occu-
pational exposure (He et al., 2009). By comparison, using a radioim-
munoassay method with a detection limit of 0.08 ng/mL, Kaddar et
al. (2009) measured BPA in 83% of the 207 randomly collected plasma
samples. Seventy‐five percent of the samples had BPA b1 ng/mL
0
10
20
30
40
50
60
70
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29
Sample No.
Concentration (ng/L)
Fig. 2. Levels of BPA (ng/L) detected in drinking water collected from household taps.
Samples 6 and 24 were from taps fitted with filter device while sample 12 was from tap
connected with PVC pipe.
0
5
10
15
20
25
123456789
25 C
50 C
Sample No.
Concentration (ng/L)
Fig. 3. Levels of BPA (ng/L) in bottled mineral water immediately after purchase (25 °C)
and after storage at 50 °C for 72 h.
Table 3
Number of samples (male and female) and percentage detected in each range of plas-
ma BPA level.
Range
(ng/mL)
Number of samples Percentage
(%)
Male Female
Not detected 38 46 83
b1.00 1 1 2
1.00–1.99 1 2 3
2.00–2.99 4 3 7
≥3.00 2 3 5
336 V.A. Santhi et al. / Science of the Total Environment 427–428 (2012) 332–338
Author's personal copy
(including below LOD) and 12% had BPA> 2 ng/mL. Similarly, 86% of
the samples in this study had no detectable level or b1 ng/mL of
BPA while only 12% had > 2 ng/mL (Table 3). Additionally in a study
by Volkel et al. (2005), all the samples had BPA at below detection
limit (0.5 ng/mL). This suggests that the level of unconjugated BPA
circulating in the blood is very low as most of the BPA is rapidly me-
tabolized by first-pass glucuronidation and eliminated in the urine.
Moreover, this finding is also consistent with the very low level of
unconjugated BPA predicted using physiologically based toxicokinetic
(PBTK) model (Edginton and Ritter, 2009).
The highest detection rate for BPA was in the b30 year age group,
followed by the age groups 30–39 years, 40–49 years and ≥50 years
(Fig. 4). This is similar to the findings by He et al. (2009) where the
highest detection rate was also in b30 age group and lowest in the
>50 age group. There is however, no significant difference in the aver-
age levels detected between males (0.41± 0.99 ng/mL) and females
(0.37± 0.91 ng/mL) although He et al. (2009) previously reported sig-
nificantly higher level in the male serum samples. The average level
detected in this study is within the range of 0.3 to 4.4 ng/mL reported
for the central measure of BPA distribution in plasma and serum sam-
ples (Vandenberg et al., 2007). However since samples for this study
was from selected communities living in the Langat River basin, the re-
sults cannot be precisely generalized to the Malaysian population.
3.6. Guideline value and exposure assessment of BPA
Currently, there is no guideline value for BPA for raw water abstract-
ed for potable use. However, the European Chemicals Bureau has de-
fined the predicted no effect concentration (PNEC) for BPA in
freshwater at 1.5 μg/L (Klecka et al., 2009). It is assumed that below
this level,aquatic organisms are protected from adverse effects from ex-
posure to BPA. Samples collected near outlets of industrial and sewage
treatment plants had levels similar to the PNEC. However, when com-
pared to the newly derived PNEC value of 0.06 μg/L by Wright-
Walters et al. (2011) which uses the weight of evidence approach
encompassing ecotoxicological endpoints of survival, growth and de-
velopment and reproduction of the most sensitive species, the detected
levels at sites S5, S6 and S7 were frequently higher. Moreover higher
levels were detected in the aquatic organisms compared to the water it-
self (Belfroid et al., 2002; Basheer et al., 2004), most likely due to bio-
accumulation. Indeed, in a recent study by Santhi et al. (2012),fish
and squid caught off the coast of Selangor had average BPA levels of
10.50 ng/g and 257.3 ng/g dry weight (d.w.), respectively.
Meanwhile, the exposure to BPA through consumption of drinking
water was assessed using the highest level detected to represent a
worst case scenario and the RfD. A daily consumption of 2 L of
water per capita for an adult Malaysian (weighing 60 kg) was used
for this purpose. The estimated daily intake (EDI) of BPA from drink-
ing water was very low (0.002 μg/kg), contributing b0.01% of the TDI.
A similar low intake was calculated by Amiridou and Voutsa (2011).
In addition, the EDI is only 0.5% of the highest daily BPA intake for
Malaysians (0.382 μg/kg), extrapolated from urinary BPA levels by
Zhang et al. (2011). By comparison, the highest EDI for seafood calcu-
lated using data from Santhi et al. (2012) is 47% suggesting that expo-
sure to BPA through drinking water is very low.
4. Conclusion
This study demonstrated that BPA is a ubiquitous contaminant in
Malaysian river water with likely source from industry and sewage
treatment plants. Low levels of BPA were detected in tap and bottled
mineral water which increased slightly in poorly stored condition. The
exposure level of unconjugated BPA in the blood plasma was low with
only 17% of the adults having detectable levels. While BPA was detected
in both the drinking water and plasma of the Langat River basin com-
munity, exposure from drinking water itself is likely to be low.
Acknowledgments
The authors would like to acknowledge the financial support from
University of Malaya (PS272/2008A), assistance from district health
personnel from Selangor during water sample collection, support
from JSPS Institutional Program for Young Researcher Overseas Visits
and Dr. Zakir Hossein for invaluable suggestions during manuscript
preparation.
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