Environ Sci Pollut Res (2014) 21:1480–1488
DOI 10.1007/s11356-013-2018-x
RESEARCH ARTICLE
Dioxin-like activity in sediments from Tai Lake,
China determined by use of the H4IIE-luc bioassay
and quantification of individual AhR agonists
Jie Xia & Guanyong Su & Xiaowei Zhang & Wei Shi &
John P. Giesy & Hongxia Yu
Received: 18 March 2013 / Accepted: 16 July 2013 / Published online: 8 August 2013
# Springer-Verlag Berlin Heidelberg 2013
Abstract Deterioration of the general ecosystem and specifically quality of the water in Tai Lake (Ch: Taihu), the third
largest freshwater in China, is of great concern. However,
knowledge on status and trends of dioxin-like compounds in
Tai Lake was limited. This study investigated AhR-mediated
potency and quantified potential aryl hydrocarbon receptor
(AhR) agonists in sediments from four regions (Meiliang Bay,
Zhushan Lake, Lake Center, Corner of Zhushan Lake, and
Meiliang Bay) of Tai Lake by use of the in vitro H4IIE-luc,
cell-based, transactivation, reporter gene assay, and instrumental analysis. Concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (Bio-TEQs) in sediments ranged from less
than the limit of detection to 114.5 pg/g, dry weight, which
indicated that organic extracts of sediments exhibited significant AhR-mediated potencies. Results of the potency balance
analysis demonstrated that acid-labile, dioxin-like compounds
represented a greater proportion of concentrations of Bio-TEQs
in sediments from Tai Lake. Concentrations of 2,3,7,8tetrachlorodibenzo-p-dioxin equivalents calculated as the sum
of the product of concentrations of individual congeners and
their respective relative potencies (Chem-TEQs) based on polycyclic aromatic hydrocarbons and/or polychlorinated biphenyls
represented no more than 10 % of the total concentrations of
Bio-TEQs.
Keywords Toxicant equivalents . In vitro bioassay .
AhR agonist . Asia
Responsible editor: Henner Hollert
Electronic supplementary material The online version of this article
(doi:10.1007/s11356-013-2018-x) contains supplementary material,
which is available to authorized users.
J. Xia : G. Su : X. Zhang (*) : W. Shi : J. P. Giesy : H. Yu (*)
State Key Laboratory of Pollution Control and Resource Reuse,
School of the Environment, Nanjing University,
Nanjing 210089, China
e-mail: zhangxw@nju.edu.cn
e-mail: yuhx@nju.edu.cn
J. P. Giesy
Department of Biomedical Veterinary Sciences and Toxicology
Centre, University of Saskatchewan, Saskatoon, SK S7N 5B3,
Canada
J. P. Giesy
Department of Biology & Chemistry, State Key Laboratory
in Marine Pollution, City University of Hong Kong,
83 Tat Chee Avenue, Kowloon, Hong Kong, SAR, China
J. P. Giesy
School of Biological Sciences, University of Hong Kong,
Hong Kong, SAR, China
Introduction
Tai Lake, the third largest freshwater lake in China, is known
for its productive fishery and natural scenery and is used as a
source of drinking water for surrounding cities, such as Wuxi
and Suzhou. In recent years, with rapid development of agriculture and industry in this area, water quality in Tai Lake has
deteriorated and is currently contaminated with both nutrients
that are causing hyper-eutrophication as well as synthetic
chemicals. Pollution of Tai Lake could threaten ecological
balance as well as health of local residents. Several studies
have focused on prevention and control of eutrophication that
has resulted in hazardous algal blooms, as well as development
of water quality criteria and ecological and human health risk
assessment of toxic contaminants in waters of the Lake (Zhou
et al. 2012; Zhang and Jiang 2005).
The aryl hydrocarbon receptor (AhR) is a liganddependent transcription factor that can be activated by
a number of chemicals that have similar structure as 2,3,7,8-
Environ Sci Pollut Res (2014) 21:1480–1488
tetrachlorodibenzo-p-dioxin (TCDD) (Hong et al. 2012a). A
range of chemicals including polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs),
polychlorinated biphenyl (PCBs), and polycyclic aromatic hydrocarbons (PAHs) can bind with and activate AhR (Van den
Berg et al. 2006; Qiao et al. 2006). When the AhR pathway is
activated by these xenobiotics, adverse effects can occur on
functions of the thyroid and immune systems and cause developmental abnormalities, hepatoxicity, and cancer in humans
and wildlife (Van den Berg et al. 2006; Hawkin and Walker
1990). Because of their hydrophobicity and persistence, these
chemicals can be accumulated and biomagnified through food
chains, and thus have the potential to adversely affect species at
higher trophic levels, including humans (Prokes et al. 2012;
Lailson-Brito et al. 2012).
There are several ways to estimate the overall AhR-mediated
potency of chemical mixtures. One approach is to sum the
products of concentrations of each measured AhR agonist
multiplied by relative potency factors (RePs or by World
Health Organization’s 2,3,7,8-tetrachlorodibenzo-p-dioxin
equivalent factors (TEFs)). The resulting overall measure of
potency is reported as chemically derived 2,3,7,8-TCDD
equivalents (Chem-TEQs; Villeneuve et al. 2002a).
However, the Chem-TEQs approach is limited by the fact that
not all AhR agonists are identified and quantified, primarily
due to a lack of suitable instrumental techniques or analytical
standards (Giesy et al. 2002; Lee et al. 2013). An alternative
approach is to determine the overall potency of a mixture of
AhR agonists by use of integrating, in vitro bioassays such as
the H4IIE-luc transactivation, reporter gene assay to measure
overall dioxin-like potency of mixtures (Schmitz et al. 1996).
Results of these assays are expressed as bioassay-derived
2,3,7,8-TCDD equivalents (Bio-TEQs). Furthermore, the
combination of in vitro bioassays and instrumental analyses
can be used to characterize both total dioxin-like activities and
known dioxin-like compounds (Koh et al. 2004). Though
pollution by dioxins and dioxin-like chemicals has been of
concern in China (Jun et al. 2004; Floehr et al. 2013), most of
the available information on Tai Lake was based on quantification of selected, individual AhR-active chemicals
(Xing et al. 2005; Mai et al. 2003). Recently, sediments
from Tai Lake were assayed for AhR-mediated EROD
induction by use of the rat hepatoma cell line; however,
only sediment from Meiliang Bay was analyzed (Qiao et al.
2006). Therefore, information on the overall potency of AhR
agonists in Tai Lake was limited.
In this study, a combination of identification and
quantification of individual AhR agonists in combination with in vitro bioassay was applied to evaluate
AhR-mediated potency of sediment extracts from four
locations in Tai Lake from May to October. The potential source and behavior of dioxin-like compounds in
Tai Lake were also characterized.
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Materials and methods
Chemicals
The 16 PAHs designated by the US Environmental Protection
Agency (including Nap, Acy, Ace, Flu, Phe, Ant, Flt, Pye,
B(a)A, Chr, B(b)f, B(a)P, InD, DBA, and B(ghi)P) as well as
28 PCBs (including PCB-8,18, 28, 52, 44, 66, 81, 77, 101, 12,
118, 114, 105, 126,153,138, 128, 167, 156, 157, 169, 187,
180, 170, 189, 195, 206, and 209) were identified and quantified. Standards were purchased from Sigma Chemical Co.
(St. Louis, MO, USA). Purities of PAHs and PCBs were 98.5
and 99.5 %, respectively.
Sampling
Samples of sediments were collected from four locations of
Tai Lake in May, June, July, September, and October of 2010
(Fig. 1). Site Y1, Y2, Y3, and Y4 were in Zhushan Lake, Lake
Center, Meiliang Bay, and the Corner of Zhushan Lake and
Meiliang Bay, respectively. Zhushan Lake and Meiliang
Bay, in the north of Tai Lake, are the most polluted
regions of Tai Lake. The Central Lake, in the center of
Tai Lake, is less affected by industrialization and urbanization. June and September are the 2 months of the
annual cycle that have the greatest precipitation.
Sediments were collected from each location into glass bottles precleaned and rinsed with methanol, dichloromethane
and hexane. Sediments were kept on ice during transportation to the laboratory and kept frozen at −20 °C
until further analysis.
Sample extraction and fractionation
Methods of extraction and fractionation were similar to those
described elsewhere with some modifications (Koh et al.
2004). Briefly, sediments were extracted by accelerated solvent extraction (Dionex ASE-350, Sunnyvale, CA, USA).
Extraction was conducted with dichloromethane at 100 °C
and pressure of 1×107 Pa. Three cycles were performed for
10 min. Extracts were treated with activated copper granules
to remove sulfur and then concentrated by rotary evaporation
to 1 ml. The extract was divided into two aliquots for use in
the bioassay or identification and quantification of individual
compounds. Raw extracts (200 μl) were passed through 10 g
of activated Florisil (60–100 mesh size; Sigma Chemical Co.,
St. Louis, MO, USA) packed in a glass column (10 mm
i.d.) for fractionation. According to the relative polarity,
three Florisil fractions (F1, F2, and F3) were collected
and used for both instrumental analyses and bioassay.
Fractions were eluted with 100 ml of hexane, 100 ml
hexane/dichloromethane (4:1), and 100 ml of methanol/
dichloromethane (1:1) (Koh et al. 2004).
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Environ Sci Pollut Res (2014) 21:1480–1488
Fig. 1 Locations of sampling
sites in Tai Lake, China
H4IIE-luc cell culture and in vitro bioassay
The H4IIE-luc, transactivation, reporter gene assay is based on
rat hepatoma cells that have been stably transfected with a
luciferase reporter gene. In this study, a high throughput 384well plate was used, which has been described previously (Su
et al. 2012). Briefly, cells were cultured in Dulbecco’s modified
eagle medium with 10 % fetal bovine serum at a maximum
density of 80–90 % confluence. Prior to confluence, cells were
trypsinized from culture dishes and then seeded into each well
of a 384-well plate at 75 μl per well. Cells were incubated for
24 h before dosing. Test wells were dosed with 0.75 μl of the
sediment extract in seven different concentrations dissolved in
dimethyl sulfoxide (DMSO; twofold dilution). Control wells
were dosed with 0.75 μl of DMSO, while the blank wells
received no treatment. Standard wells were dosed with 10
different known concentrations of TCDD. Three replicates were
conducted per treatment, including TCDD standards. On the
fifth day, cells were lysed and luciferase activity was determined
by a commercial kit (Promega Corporation, Madison, WI,
USA) in a microplate reader (BioTek Instruments Inc.,
Winooski, VT, USA). Responses to AhR agonists were quantified by measuring the relative luminescence units and were
converted to percentages of the mean maximum response observed for the TCDD standard curve (%-TCDD-max). BioTEQs were determined by use of dose–response curves generated by testing samples at multiple (at least three points) of
dilutions (Villeneuve et al. 2000a) and comparing the magnitude of response to that of 2,3,7,8-TCDD. Labile chemicals
(including PAHs) can be removed by treatment with concentrated
sulfuric acid, leaving only persistent PCBs, OCPs, and potentially PCDD/PCDFs (Blaha et al. 2006; Hilscherova et al.
2000). The removal efficiency of 16 PAHs ranged from 95
to 100 % (Electronic supplementary material (ESM)
Table S1). Stability of PCBs and TCDD with concentrated
sulfur acid was also evaluated with H4IIE-luc assay. PCBs
and TCDD would not be degraded after treatment with
concentrated sulfuric acid (ESM Fig. S1).
Instrumental analysis
The 16 indicator PAHs and 28 PCB congeners were identified
and quantified by use of a Thermo Scientific TSQ Quantum
GC (Waltham, MA, USA), coupled with an Agilent DB-XLB
column (15 m×0.25 mm×0.25 μm, USA). The mass spectrometer detector was operated in electron impact ionization
mode. Samples and standards were analyzed in selected reaction monitoring (SRM) mode. Quantification and qualification were processed by SRM modes. The precursor ion and
product ions selected in SRM mode for each chemical were
based on the mass spectrum of the standard solution. Detailed
information about precursor ion, product ions, ions ratio, and
collision energy are given in ESM (Table S2). Detection limits
for PAHs and PCBs ranged from 0.01 to 0.1 and 0.003 to
0.03 ng/g, respectively.
Data analysis
The Z factor was used to evaluate the performance of the 384well format H4IIE-luc assay for in high-throughput screening
Environ Sci Pollut Res (2014) 21:1480–1488
1483
of environmental samples. The Z factor is defined in terms of
four parameters: the means (μp and μn) and standard deviations (σp and σn) of both the positive and negative controls
(Sui and Wu 2007; Eq. 1).
3 σp þ σn
Z factor ¼ 1− ð1Þ
μp þ μn Due to small responses, Bio-TEQs were expressed
by the ratio of the EC20 (20 % effect concentration)
values for each sediment sample to that of the positive
control TCDD (Wolz et al. 2008). Bio-TEQs were
calculated as mean values of n = 2 individual H4IIE-luc
bioassay (Eq. 2).
Bio−TEQs ¼ EC20;TCDD =EC20;sample
ð2Þ
TCDD equivalents based on Chem-TEQs were calculated
as the sum of Chem-TEQs contributed by individual
chemicals as the sum of the products of concentrations of
individual AhR-active chemicals by their respective TEFs or
RePs (Su et al. 2012). Seven dioxin-like PAHs (BaA, Chr,
BbF, BkF, BaP, IcdP, and DbahA) and 12 dioxin-like PCBs
(PCB-77, 81,105,114, 118, 123, 126, 156, 157, 167, 169, and
189) were used to calculate the concentrations of PAH-TEQ
and PCB-TEQ as previously described (Villeneuve et al.
2002b; Nakata et al. 2003; Van den Berg et al. 1998). ChemTEQs for each sample were calculated (Eq. 3).
Xn
Chem –TEQs ¼
concentrationi TEFi =REPi
ð3Þ
i¼1
Spearman rank correlations were used to examine the
strength of association between different data sets, including time trends of chemical concentration for each
sampling location and bioassay activity among different
sites. Concentrations of analytes in sediments are presented as
mathematic means. Dose–response curves of TCDD and sediment samples in the bioassay were calculated with software
Graphpad Prism 4.0 (GraphPad, San Diego, CA, USA) using
four parameters sigmoidal model. Other figures were generated with OriginPro 8 (OriginLab Corporation, Northampton,
MA, USA).
Results
H4IIE-luc assay validation
A 384-well plate format for the H4IIE-luc cell reporter gene
assay showed a strong concentration-dependent response to a
serial concentration of standard chemical TCDD (Fig. 2).
Fig. 2 Concentration-dependent responses and a fitted curve for TCDD
in H4IIE-luc cell reporter gene assay (mean values and standard deviations were plotted along with the fitted curve)
Significant responses (2.4 %-TCDD-max) were defined as
those resulting in a response three times as great as the
standard deviation of the mean solvent control responses,
which were also regarded as the detection limit for the bioassay. The EC20, EC50, and EC80 for luciferase induction by
TCDD were determined to be 2.97±0.78, 14.26±1.97, and
68.42±15.03 pM, respectively. The Z factor for the 384-well
format H4IIE-luc assay was calculated to be 0.86 (when a Z
factor is between 0.5 and 1.0, the developed high-throughput
assay was judged to be excellent). The results indicated that
the H4IIE-luc assay employed here was sufficiently sensitive
to provide quantitative results.
Dioxin-like activity of the raw extracts
Extracts of sediments from all four locations in Tai Lake from
May to October induced AhR-mediated potency in the H4IIEluc bioassay (Table 1). Magnitudes of responses caused by
unfractionated extracts of sediments were less than the maximum response produced by a 1,500 pM TCDD standard and
ranged from 8 to 32 %-TCDD-max. EC20 values were
used to calculate Bio-TEQs in extracts. Concentrations of
Bio-TEQs in sediments ranged from less than the limit of
detection to 114.5 pg TCDD/g, dry weight (dw). The maximum responses and Bio-TEQs for different sites were significantly different. The rank-order of dioxin-like potency, based
on %-TCDD-max and Bio-TEQs, was Y1 (mean, 27 %;
78.5 pg/g, dw)>Y3 (mean, 18 %; 21.4 pg/g, dw)>Y2 (mean,
12 %; 2.5 pg/g, dw)>Y4 (mean, 12 %; 1.9 pg/g, dw). Six
sediments unfractionated or acidified extracts of Y1 and Y3,
with responses above 20 %-TCDD-max, were further selected
to determine the main AhR agonists.
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Table 1 Concentrations of Bio-TEQs (in picogram per gram) in sediments of Tai Lake, Jiangsu Province, China
Y1
Y2
Y3
Y4
Location
%-TCDD-max
Bio-TEQ
May
June
July
September
29±3
17±5
32±1
25±6
93.6±10.2
13.5±2.0
114.5±15.6
57.4±9.2
October
May
June
July
September
October
May
June
July
September
October
May
June
July
September
October
32±4
9±3
12±1
12±1
11±3
16±1
20±2
14±2
31±6
14±2
12±4
8±2
14±2
14±3
10±2
14±2
113.6±15.9
NC
1.5±1.1
2.4±1.5
1.0±0.6
7.4±2.9
17.5±2.1
5.0±1.2
76.9±12.3
5.3±1.2
2.3±1.8
NC
4.1±1.3
3.1±1.6
NC
2.1±1.2
NC not calculated
AhR-mediated activity of fractions
Three Florisil fractions of the six selected extracts were analyzed to examine the bioassay response to specific classes of
compounds separated based on polarity. F1 samples caused
the least response of the H4IIE-luc response (Fig. 3). The ratio
of AhR potency of a fraction to that of total raw extract (F/T)
Fig. 3 Emission of light by
luciferase in the H4IIE-luc cell
bioassay elicited by sediment
fractions (F1, F2, F3) and acidtreated extracts. Response
presented as percentage of the
maximum response observed for
2,3,7,8-tetrachlorodibenzo-pdioxin standard
for F1 was 7–18 %-TCDD-max (mean, 13 %). PCBs and few
PAHs were detected in F1. Among the six sediment samples,
PCBs were detected in Y1-May (2.78×10−1 ng/g, dw) and
Y3-May (6.50×10−1 ng/g, dw). PCB-TEQ of Y1-May and
Y3-May were 2.77×10−1 and 1.46×10−3 pg/g (Table 2). F2
samples induced moderate responses (F/T ratio, 11–86 %TCDD-max with a mean 42 %) in the H4IIE-luc bioassay
(Fig. 3). Among the six sediment samples, all 16 target PAHs
were detected in F2 at concentrations ranging from 7.74×101
to 5.11×102 ng/g, dw. This was consistent with the previous
publications (Koh et al. 2004). PAH-TEQ of the six sediment
samples ranged from 1.05×10−1 to 2.94 pg/g (Table 2).
Among the three fractions tested, F3 induced the
greatest magnitude of response (F/T ratio, 60–99 %TCDD-max with a mean of 77 %) in the H4IIE-luc
bioassay (Fig. 3). PAHs and PCBs were not detected
in F3. Previous studies have reported that PCDD/PCDFs
partition primarily to F2, but some may carry over into
F3 (Koh et al. 2004). The six raw extracts of sediments
were treated with concentrated sulfuric acid in order to
determine whether the AhR activity was due to acidlabile substances or PCDD/PCDFs.
AhR potency of total acid-treated extracts
Acid-labile compounds were unstable to oxidative breakdown
and are removed by concentrated sulfuric acid and acid-stable
compounds were effectively isolated from raw extracts
(Woelz et al. 2011). After treatment with concentrated sulfuric
acid, TCDD-max values of the six extracts ranged from 6 to
10 % (Fig. 3). The ratio of AhR potency of acid-treated
extracts to that of respective total untreated extracts were from
22 to 38 % (mean, 29 %). Due to the small TCDD-max values,
which were less than the linear range of the dose–response
Environ Sci Pollut Res (2014) 21:1480–1488
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Table 2 Concentrations of Chem-TEQs based on PAHs and PCBs in sediments of Tai Lake, Jiangsu Province, China
Chemical
TEF/REPa
B[a]A
Chr
B[b]F
B[k]F
B[a]P
I[cd]P
D[ah]A
PAH-TEQ
PCB77
PCB81
PCB105
PCB114
PCB118
PCB123
1.90×10−6
2.30×10−6
5.10×10−6
1.40×10−4
1.60×10−6
1.50×10−5
4.60×10−6
PCB126
PCB156
PCB157
PCB167
PCB169
PCB189
PCB-TEQ
Chem-TEQs
Bio-TEQs
1.00×10−1
3.00×10−5
3.00×10−5
3.00×10−5
3.00×10−2
3.00×10−5
1.00×10−4
3.00×10−4
3.00×10−5
3.00×10−5
3.00×10−5
3.00×10−5
Concentrations at sampling sites (in picogram per gram, dw)
Y1-May
Y1-Jul
Y1-Sep
Y1-Oct
Y3-May
Y3-Jul
ND
2.72×104
2.33×104
6.00×102
4.80×103
3.40×103
1.40×103
3.31×10−1
2.18×101
8.78×10−1
1.97
1.27
ND
4.81×10−1
1.63×104
2.73×104
2.06×104
4.20×103
8.10×103
4.00×102
ND
8.06×10−1
ND
ND
ND
ND
ND
ND
4.62×104
7.39×104
6.07×104
1.53×104
3.09×104
1.19×104
ND
2.94
ND
ND
ND
ND
ND
ND
4.05×103
1.22×104
1.68×104
5.60×103
6.50×103
1.13×104
7.00×102
1.09
ND
ND
ND
ND
ND
ND
6.60×103
1.36×104
1.32×104
ND
7.00×102
2.30×103
8.00×102
1.50×10−1
2.44
5.47×10−1
2.24
ND
1.72×101
1.52×101
2.02×104
3.21×104
2.42×104
7.10×103
1.84×104
6.30×103
ND
1.35
ND
ND
ND
ND
ND
ND
2.75
ND
1.38
ND
ND
ND
2.77×10−1
6.08×10−1
93.6
ND
ND
ND
ND
ND
ND
ND
8.06×10−1
114.5
ND
ND
ND
ND
ND
ND
ND
2.94
57.4
ND
ND
ND
ND
ND
ND
ND
1.09
113.6
ND
1.22×10−1
ND
4.70×10−1
ND
ND
1.46×10−3
1.52×10−1
17.5
ND
ND
ND
ND
ND
ND
ND
1.35
76.9
ND not detected
a
TEF/REP values were obtained from Villeneuve et al. (2002b), Nakata et al. (2003), and Van den Berg et al. (1998)
curve by TCDD, Bio-TEQs in extracts after treatment with
sulfuric acid were not calculated.
Discussion
All tested sediment extracts from Tai Lake exhibited dioxin-like
activity in the H4IIE-luc bioassay. Dioxin-like activities of
sediments from different sites were significantly different.
Concentration of Bio-TEQs in extracts from Y1 (13.5–
114.5 pg/g) and Y3 (2.3–76.9 pg/g) were substantially greater
than Y2 (NC to 2.4 pg/g) and Y4 (NC to 4.1 pg/g), which
illustrated that pollution at sites Y1 and Y3 was more serious
than Y2 and Y4. The cities of Yixing, Changzhou, and Wuxi,
which are adjacent to locations Y1 and Y3, where the greatest
Bio-TEQ was observed, are characterized by their rapid industrial and urban development. Multiple industries, such as machinery works, paper mills, chemical plants, and electroplating
factories are located at these cities. Effluents and runoff from
factories and cities are likely sources of Bio-TEQs in sediments
at locations Y1 and Y3. Locations Y2 and Y4, in the center of
Tai Lake, are less affected by environments around Tai Lake.
Concentrations of Bio-TEQs in sediments collected in June and
September were lesser than those collected during other
months. This could be due to dilution by rainfall. In Tai Lake,
the rainfall in June and September is more than in any other
months, which could dilute the pollutants in the aquatic environment. The screening results of raw extracts of sediments
from Tai Lake indicated that total potencies of AhR agonists
were greater than those of sediments from the north coast of the
Bohai Sea (<ND–28 pg/g, dw) and Kwangyang Bay, USA
(ND–6.44, dw) (Hong et al. 2012a; Koh et al. 2005), similar
to those of sediments from Osaka Bay, Japan (3.7–140 pg/g,
dw; Takigami et al. 2005), but lesser than those of sediments
from the Haihe, China (330–930 pg/g, dw), Dagu, China
(1,200–13,900 pg/g, dw), Hyeongsan Rivers, South Korea
(14–1,500 pg/g, dw), and Saginaw River, USA (0.01–
3,630 pg/g, dw; Koh et al. 2004; Song et al. 2006; Kannan
et al. 2008; Table 3).
Total concentrations of the 16 indicator PAHs in sediments
from Tai Lake ranged from 7.44×10−1 to 5.11×102 ng/g, dw,
with a mean of 2.20×102 ng/g, dw. Concentration of PAHs in
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Environ Sci Pollut Res (2014) 21:1480–1488
Table 3 Comparisons of concentrations of Bio-TEQs and Chem-TEQs in sediments from Tai Lake determined in this study with those reported
previously
Country
Location
Bio-TEQs
Chem-TEQs
References
PCDD/PCDFs-TEQ
PCBs-TEQ
PAHs-TEQ
China
Meiliang Bay of Tai Lake
17.8–35.8
1.9–6.0
0.2–0.4
14.5–33.8
Qiao et al. (2006)
330–930
1,200–13,900
8.5–336.0
ND–28
17.45–114.50
3.7–140
6.76–19.13
21.93–892.59
6.2–12.3
ND–3.4
NA
1.8–48
9.4–37.7
NA
ND–0.28
0.04–44
NA
NA
4.9–20.5
0.27–9.3
0.15–2.94
NA
Song et al. (2006)
Japan
Haihe River
Dagu River
Wenyu River
North coast of Bohai Sea
Tai Lake
Osaka Bay
South Korea
West coast area
Hyeongsan River
Lake Shiwa
Masan Bay
Kwangyang Bay
Taean area
ND–11
14–1,500
3.51–327
2.3–284
NA–6.44
1.6–2,500
ND–1.3
5.5–1,000
NA
NA
NA
NA
NA
0.16–3.7
0.78–42.0
0.19–77.7
ND–3.83
NA
0.17–0.93
0.20–88
0.01–3.49
0.54–3.64
0.02–1.10
0.01–2.6
Hong et al. (2012a)
Koh et al. (2004)
Koh et al. (2005)
USA
Saginaw River
Shiawassee River
Saginaw Bay
Danube River
Tiete River
0.01–3,630
0.01–3,100
ND-547
110–1,400
ND-24169.73
3.0–3,820
0.2–9.8
3.0–266
28–130
NA
Kannan et al. (2008)
NA
NA
NA
NA
NA
9.10–567.05
Germany
Brazil
Luo et al. (2009)
Hong et al. (2012a)
This study
Takigami et al. (2005)
Hong et al. (2012c)
Keiter et al. (2008)
Suares Rocha et al. (2010)
NA not analyzed, ND not detected
Tai Lake, observed in this study, were lesser than those reported
for sediments collected several years ago (Qiao et al. 2006;
Zhang et al. 2011; Tao et al. 2010), which indicated that
concentrations of PAHs might be decreasing because of pollution control measures undertaken during the past several years.
Using ratios between individual PAHs as a forensic tool
(Baumard et al. 1999; Dickhut et al. 2000; Soclo et al. 2000),
the primary sources of the PAHs observed in sediments were
attributable to a mixture of combustion of fossil fuels
(pyrogenic) and discharge of petroleum-derived materials
(petrogenic; Table 3). Thus, to control inputs of PAHs would
require controlling different sources in Tai Lake. In general,
neither individual PAHs concentrations nor total concentrations
of PAHs exceeded respective effect range low values (Long
et al. 1995). Seven dioxin-like (DL-PAHs) including BaA, Chr,
BbF, BkF, BaP, IcdP, and DBahA were detected in most
Table 4 Characteristic ratios of
PAHs derived from several
sources
NA not available
a,b
From Baumard et al. (1999),
Dickhut et al. (2000), Soclo et al.
(2000)
sediments in Tai Lake (Table 2). No significant differences
were observed among concentrations of DL-PAHs in extracts.
Among these DL-PAHs, concentrations of Chr and BbF were
greatest in Tai Lake, contributing 32 and 27 % of the total
concentrations of PAHs, respectively. Chr and BbF are generated by coal combustion and diesel-powered vehicles, respectively (Larsen and Baker 2003). Power plants around Tai Lake
and fishing boats in Tai Lake are likely the main sources of
Chr and BbF, respectively (Table 4).
Concentrations of PCBs in Tai Lake were relatively small.
Among the six sampling locations, PCBs were detected only
in sediments from Y1-May and Y3-May. Among the 12
dioxin-like PCBs (DL-PCBs), PCB-126 with the greatest
toxic potency, was detected in sediments from only Y1
(May), with a concentration of 2.75×10−3 ng/g, dw. Total
concentrations of PCBs in sediments from Y1 (May) and Y3
PAH source
B[a]A/Chry
Phe/An
Flu/Pyr
Flu/(Flu+Pyr)
LMW/HMW
Pyrolytic origina
Petrogenic origina
Automobilesb
>1
<1
0.53±0.06
<10
>15
NA
>1
<1
NA
>0.5
<0.5
NA
Low
High
NA
Coal/cokeb
Woodb
Smeltersb
This study
1.11±0.06
0.79±0.13
0.60±0.06
0–0.63
NA
NA
NA
1.27–34.78
NA
NA
NA
1.32–7.90
NA
NA
NA
0.57–0.89
NA
NA
NA
0.1–0.5
Environ Sci Pollut Res (2014) 21:1480–1488
(May) were lesser than those reported in other publications
(Zhang and Jiang 2005; Qiao et al. 2006).
Because Chem-TEQs, based on PAHs and PCBs,
accounted for less than 10 % of the Bio-TEQs (Table 2) and
acid-treated extracts accounted for 22–38 % of TCDD-max of
the raw sediment extracts, PCDD/PCDFs contributed less than
38 % of TCDD-max of the raw extracts. Because concentrations of Bio-TEQs exceeded those of Chem-TEQs,
unidentified AhR agonists occurred in sediments of Tai Lake.
Some compounds such as polybrominated biphenyls, chlorinated and brominated polycyclic aromatic hydrocarbons,
polychlorinated naphthalenes, hydroxylated polybrominated
biphenyls ethers, methoxylated polybrominated biphenyls
ethers, acid-labile compounds, amd humic and fulvic acids
have been reported to have dioxin-like activity (Su et al.
2012; Bittner et al. 2009, 2011; Chen and Bunce 2003;
Villeneuve et al. 2000b; Luo et al. 2009; Hong et al. 2012b;
Horii et al. 2009). In our study, concentrated sulfur acid was
used to remove acid-labile compounds in order to qualitatively
classify AhR mediated potency of PCDD/PCDFs in raw
extracts. The ratio of AhR potency of acid treated extracts to
that of total raw extract were determined to be from 22 to 38 %
(mean, 29 %). Among the three fractions tested, the greatest
induction of luciferase activity was caused by F2 and F3.
These results suggest that acid-labile, moderate polar, and
polar compounds were the primary AhR agonists contributing
to Bio-TEQs in sediments of Tai Lake.
Acknowledgments The research was supported by the Environmental
Protection Public Welfare Scientific Research Project, State Environmental
Protection Administration People’s Republic of China (grant no.
201209016), Jiangsu Provincial Key Technology R&D Program
(#BE2011776). Prof. Giesy was supported by the program of 2012 "High
Level Foreign Experts" (#GDW20123200120) funded by the State Administration of Foreign Experts Affairs, China as well as the Canada Research
Chair program, an at large Chair Professorship at the Department of Biology
and Chemistry and State Key Laboratory in Marine Pollution, City University of Hong Kong, the Einstein Professor Program of the Chinese Academy of Sciences. Prof. Zhang was supported by a Program of the Ministry
of Education of China for New Century Excellent Talents in Universities.
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