Full text of DOI:10.1002/etc.5620210906

Environmental Toxicology and Chemistry, Vol. 21, No. 9, pp. 1796–1803, 2002
2002 SETAC
Printed in the USA
730-7268/02 $9.00 .00
᭧
0
ϩ
ANALYSIS OF TRACE ORGANIC CONTAMINANTS IN SEDIMENT, PORE WATER,
AND WATER SAMPLES FROM ONSAN BAY, KOREA: INSTRUMENTAL ANALYSIS
AND IN VITRO GENE EXPRESSION ASSAY
C
HUL-HWAN KOH,*† JONG SEONG KHIM,†‡ DANIEL L. VILLENEUVE,‡ KURUNTHACHALAM KANNAN,‡ and
J
OHN P. G IESY‡
†
School of Earth and Environmental Sciences (Oceanography), College of Natural Sciences, Seoul National University,
Seoul 151-742, South Korea
‡
National Food Safety and Toxicology Center, Department of Zoology, and Institute for Environmental Toxicology,
Michigan State University, East Lansing, Michigan 48824-1311, USA
(
Received 24 October 2001; Accepted 7 March 2002)
Abstract—Persistent organic pollutants and alkylphenols (APs) were determined in sediment and water samples from Onsan Bay,
Korea, by using instrumental analysis and in vitro gene expression cell bioassay. Polycyclic aromatic hydrocabons (PAHs) were
the predominant compounds in sediments with concentrations as great as 573 ng/g dry weight. The PAH concentrations were
elevated in sediment from inland rivers that flow through Onsan City (mean: 116 ng/g dry wt) and discharge into Onsan Bay.
Concentrations of polychlorinated biphenyls (PCBs) in sediments ranged from
organochlorine (OC) pesticides analyzed (hexachlorobenzene, hexachlorocyclohexanes, chlordanes, and DDTs), DDT concentrations
were the greatest, ranging from 0.01 to 7.58 ng/g dry weight. The spatial gradient of contaminant concentrations suggested that
Ͻ1.00 to 56.2 ng/g dry weight. Among different
Ͻ
streams and rivers are the major sources of PCBs, PAHs, and APs to the bay. Maximum concentrations of nonylphenol, octylphenol,
and bisphenol A in sediments were 860, 11, and 204 ng/g dry weight, respectively. Screening of Onsan Bay sediment samples for
dioxinlike activity with the H4IIE-luc in vitro cell bioassay revealed that 17 of 22 samples contained significant dioxinlike activity.
Further fractionation of sediment extracts indicated that mid-polar and more polar fractions were responsible for the significant
dioxinlike activity. Based on a mass balance analysis, PAHs apparently accounted for only a small portion of dioxinlike responses
elicited by sediment extracts. Only one raw extract of sediment elicited a significant estrogenic response by MVLN cells. The
combination of instrumental analysis and in vitro bioassay was useful to assess sediment quality and characterize the causative
agents or potential toxic compounds present.
Keywords—Sediment
Persistent organic pollutants
Polycyclic aromatic hydrocarbons
Xenoestrogen
In vitro
bioassay
INTRODUCTION
Despite the potential for direct and accidental releases of
organic contaminants into Onsan Bay, little was known about
contamination in this region. This study represents one of the
first efforts to examine the concentrations, distribution, and
biological potency of organic contaminants present in Onsan
Bay sediments and waters. The compounds examined in this
study were polycyclic aromatic hydrocarbons (PAHs), poly-
chlorinated biphenyls (PCBs), organochlorine (OC) pesticides,
nonylphenol (NP), octylphenol (OP), and bisphenol A (BPA).
The PAHs are derived from not only anthropogenic sources
such as waste incineration, coal gasification, and accidental
oil spills, but also are combustion products of fossil fuels and
wood [1]. Because of their mutagenicity, carcinogenicity, and
toxic effects similar to that of dioxins, their concentrations
have been evaluated in a variety of environmental matrices
including sediment [2–4]. Polycyclic aromatic hydrocarbons
have been identified as prominent contaminants in sediments
from several Korean coasts [5–7].
Onsan Bay, located on the east coast of Korea, contains
several industrial complexes and a large commercial harbor.
Industrial complexes contain several heavy metal (zinc, alu-
minum, and copper) and petrochemical industries, paper mills,
and the largest nonferrous metal industry in Korea. Heavy
industrial and shipping activities around the Onsan Bay region
and rapid urbanization of Onsan City have resulted in envi-
ronmental contamination since the 1980s. Approximately
3
1
40,000 t of industrial and domestic wastes from more than
00 industrial complexes are discharged daily into the bay
through Woihwang River and Daejeong Stream (Fig. 1). About
one half (150,000 t) of the wastes receive primary sewage
treatment, whereas the rest are discharged directly into the
bay. Pollution in this bay has been suspected to be related to
Onsan disease among inhabitants who consumed seafood col-
lected from the bay in 1985. The disease is characterized by
nausea, skin discoloration, and in extreme cases, death. The
Korean government planned to relocate 27,000 inhabitants
who lived around the bay to other areas because the bay was
a serious environmental and health problem. Heavy metals
such as cadmium were suspected contaminants for Onsan dis-
ease; however, the actual cause of the disease has never been
determined.
Recently, concern has increased about contamination by
halogenated aromatic hydrocarbons, such as DDTs, PCBs, po-
lychlorinated dibenzo-p-dioxins, and polychlorinated diben-
zofurans in Korea [5–11]. Although sources, emission, trans-
port, fate, and effects of persistent organic pollutants have long
been studied worldwide, little is known about these compounds
in Korea because of the lack of analytical facilities.
*
To whom correspondence may be addressed (kohch@snu.ac.kr).
Alkylphenols (APs) such as NP and OP are another class
1
796

Trace organic contaminants in aqueous media
Environ. Toxicol. Chem. 21, 2002
1797
distribution in the outer bay (Onsan Bay; ON, OS, and O
locations). Whole sediment samples (three replicates) were
collected from 22 locations except for one (D5) by a Van Veen
grab sampler (25
ϫ 40 ϫ 30 cm) and surface sediment (0–5
cm) samples subcollected for chemical analysis and bioassay.
Samples were homogenized in precleaned aluminum contain-
ers and pebbles and twigs were removed. Samples were stored
in precleaned amber glass bottles (250 ml) and transported on
dry ice to the laboratory. Samples were freeze-dried and
ground with a mortar and pestle and stored in precleaned high-
density polyethylene bottles at
Ϫ20ЊC until extraction. Total
organic carbon was analyzed with a CHN analyzer [6]. Surface
water samples (about 16 L) were collected from 10 locations
(
W0, W1, W2, D2, D4, D5, ON1, OS1, O4, and O10) in
precleaned 4-L amber glass bottles and stored on ice. Eleven
pore-water samples (about 1 L) were obtained from corre-
sponding sediment samples (W0, W1, W2, D1, D2, D3, D4,
ON1, OS1, O4, and O10) by using a pressurized (squeezing)
method immediately after sampling surface sediment [16]. Wa-
ter and pore-water samples were stored in 4-L amber glass
bottles and transported on ice to laboratory. Water samples
were further separated into particulate matter and dissolved
fraction by using glass fiber filters and membrane filters (final
pore size: 0.45
␮m). Particulate matter samples were freeze-
dried and stored in precleaned aluminum foil at
extraction.
Ϫ20ЊC until
Fig. 1. Map of the study area. Samples (22 sediment, 10 water, and
1
1 pore water) were collected from 23 locations from Woihang River
(
(
W0–2 locations), Daejeong Stream (D0–6 locations), and Onsan Bay
ON1–3, OS1–3, and O4–10 locations), Korea, in April 1999.
Sample preparation
Polycyclic aromatic hydrocarbons, PCBs, OC pesticides
DDTs, hexachlorocyclohexanes, chlordanes, and hexachlo-
robenzene), NP, OP, and BPA were analyzed by previously
described methods [5]. Sediment and particulate matter sam-
ples were Soxhlet extracted for 20 h with dichloromethane
Burdick & Jackson, Muskegon, MI, USA). Solid-phase sam-
ples were treated with activated copper granules to remove
residual sulfur from the extracts. Aqueous phase samples such
(
of contaminants found frequently in sediments from Korea
5,6]. Alkylphenols are degradation products of AP ethoxy-
[
lates, which are nonionic surfactants widely used for industrial
and household purposes [12]. Bisphenol A is another envi-
ronmental contaminant released through its use in polycar-
bonate plastics, epoxyresins, and phenoxy resins, which are
utilized in food storage containers and in dental sealants [5].
These compounds enter the aquatic environment via industrial
and municipal wastewater effluents. Information on their dis-
tribution in sediments is necessary to understand their effects
on sediment bound organisms and to elucidate their environ-
mental fate. Nonylphenol, OP, and BPA were analyzed in this
study.
Because of the complex nature of contaminants in sedi-
ments, sample extracts were fractionated according to polarity
to isolate and identify target contaminants. Both instrumental
analyses and in vitro recombinant cell bioassays that used
H4IIE-luc cells for dioxinlike activity and MCF-7-luc (MVLN)
cells for estrogenic activity were performed to quantify target
contaminants and to evaluate dioxinlike and estrogenic poten-
cies [5,13]. Assessment based solely on instrumental analysis
may over or underestimate potential hazards of sediment con-
tamination. Thus, a combination of instrumental analysis and
in vitro bioassay were used to assess sediment contamination
(
as water and pore water were passed through Empore
௢ solid-
phase disks (3M, St. Paul, MN, USA) and then extracted with
hexane and dichloromethane. Raw extracts (REs) were con-
centrated and the volume was adjusted to 2 ml in hexane.
One milliliter of RE was used for in vitro bioassay. The
remaining 1 ml was passed through a 10-g activated Florisil
column (60–100 mesh size; Sigma, St. Louis, MO, USA) for
further fractionation. Three fractions (F1, F2, and F3) were
collected according to polarity and used for both instrumental
analyses and bioassay. Florisil separation was confirmed by
using a spike recovery test and standard reference material,
1
974a sediment (National Institute of Science and Technology,
Gaithersburg, MD, USA). Recoveries of target analytes
through the analytical procedures were between 90% and
1
05%. Procedural blanks were analyzed with every set of six
samples to check for interference or contamination arising
from solvents or glassware.
[
13–15].
Instrumental analysis
The PAHs were quantified by using a Hewlett-Packard 5890
series II gas chromatograph equipped with a 5972 series mass
spectrometer detector (Avondale, PA, USA). Organochlorine
pesticides and PCBs were quantified by using a gas chro-
matograph (Perkin-Elmer series 600, Norwalk, CT, USA)
equipped with 63Ni electron-capture detector. An equivalent
mixture of 98 individual PCB congeners (AccuStandard, New
Haven, CT, USA) and a mixture of OC pesticides (CLP-023R
MATERIALS AND METHODS
Sampling
Sediment and water samples were collected from 23 lo-
cations in Onsan Bay during April 1999 (Fig. 1). Sampling
was designed to determine potential sources of contaminants
from inland regions such as the Woihwang River and Daejeong
Stream (designated as W and D locations) and to elucidate the

1
798
Environ. Toxicol. Chem. 21, 2002
Table 1. Concentrations (ng/g dry wt) of target organic compounds in sediments from Onsan Bay, Koreaa
C.-H. Koh et al.
HCB
Region
Location
PAHs
NP
OP
BPA
PCBs
DDTs
CHLs
HCHs
Woihwang River
W0
W1
W2
D0
D1
D2
D3
D4
D6
ON1
ON2
ON3
OS1
OS2
OS3
O4
O5
O6
O7
O8
Ͻ
10.0
77.0
Ͻ
20.8
1.00
Ͻ
1.00
1.00
1.00
1.00
Ͻ
Ͻ
1.00
1.00
1.59
1.00
6.87
10.1
5.47
0.07
0.02
0.35
7.55
7.58
0.02
1.20
1.67
3.06
2.54
0.01
0.47
0.61
0.76
0.20
0.25
0.33
0.05
0.06
0.05
0.03
0.15
Ͻ
Ͻ
0.01
0.01
0.04
0.01
0.95
0.01
0.13
0.81
0.01
0.08
0.01
0.01
0.01
0.01
0.03
0.05
0.05
0.01
0.01
0.01
0.01
0.01
0.02
1.02
3.37
0.04
0.79
0.02
0.26
3.17
0.78
1.12
0.01
0.02
0.76
0.02
0.68
0.26
0.44
0.10
0.02
0.12
0.02
0.41
0.03
0.69
0.55
0.03
NA
1.69
2.58
4.44
0.21
NA
Ͻ
Ͻ
Ͻ
11.01
Ͻ
573
Ͻ
113
Ͻ
4.78
6.23
Daejeong Stream
Onsan Bay
10.0
Ͻ
204
Ͻ
Ͻ
1.00
NA
Ͻ
Ͻ
860
14.4
100
96.4
10.0
34.5
1.00
1.39
5.75
1.00
1.00
1.00
1.00
1.00
1.00
1.00
Ͻ
1.00
1.21
8.93
1.00
1.00
1.00
1.00
1.00
1.00
19.6
56.2
3.33
214
21.4
10.0
Ͻ
1.00
1.86
1.26
1.00
3.46
1.39
NA
1.00
1.00
1.00
1.00
2.68
3.80
1.00
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
NA
7.79
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
NA
j
NA
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
1.57
0.08
NA
38.7
35.9
19.3
17.9
41.6
27.0
10.0
10.0
10.4
27.7
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
0.01
0.15
0.20
0.05
0.05
0.04
0.13
NA
NA
NA
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
4.33
1.00
1.00
1.00
1.00
1.00
1.00
1.00
Ͻ
1.00
1.00
1.00
1.00
1.00
1.00
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
O9
O10
Ͻ
Ͻ
1.00
1.00
0.04
0.01
Ͻ
Ͻ
a
PAHs
polychlorinated biphenyls, sum of 98 individual congeners; DDTs
dichlorodiphenyldichloroethylene; CHLs sum of and -chlordanes; HCHs
hexachlorobenzene; NA not analyzed.
ϭ
polycyclic aromatic hydrocarbons, sum of 16 priority components; NP
sum of p,p
sum of
ϭ
nonylphenol; OP
-DDT, p,p
-,
ϭ
octylphenol; BPA
-dichlorodiphenyldichloroethane, and p,p
-, and -hexachlorocyclohexanes; HCB ϭ
ϭ bisphenol A; PCBs
ϭ
ϭ
Ј
Ј
␤
Ј
ϭ
␣
␥
ϭ
␣
␥
ϭ
and CLP-024R, AccuStandard) were used as a standard [17].
Reverse-phase high-performance liquid chromatography with
fluorescence detection was used to quantify NP and OP. Further
details of sample preparation and instrumental analysis are
described elsewhere [5,18].
RESULTS AND DISCUSSION
Concentration and distribution of persistent
organic pollutants
The relative abundance of organic residues in sediment was
in the order PAHs APs BPA PCBs OC pesticides
Table 1). The PAHs were detected in 14 of 20 sediment sam-
ഠ
Ͼ
Ͼ
Ͼ
(
In vitro bioassay
ples analyzed (Table 1). The PAH concentrations were as great
as 573 and 214 ng/g dry weight in sediment from locations
W2 and D4, which correspond to Woihwang River and Dae-
jeong Stream, respectively. Mean PAH concentrations in sed-
iments from the Woihwang River (mean: 218 ng/g dry wt) and
Daejeong Stream (mean: 65.4 ng/g dry wt) locations were
approximately 10- and threefold greater than those from outer
Onsan Bay sites (mean: 19.2 ng/g dry wt), respectively. No
spatial gradient was found in concentrations of PAHs in river
or stream sediment. This suggests localized inputs of PAHs
from sources along the river and stream. Four-ring aromatic
hydrocarbons, such as fluoranthene and pyrene, were the pre-
dominant PAHs in Onsan Bay sediment. Molecular ratios of
specific PAH compounds, such as fluoranthene to pyrene ratio
and indeno[1,2,3-cd]pyrene to benzo[ghi]perylene ratio, were
calculated to evaluate the potential sources of PAHs [3]. The
fluoranthene to pyrene ratios for Onsan Bay sediment samples
varied depending on the locations with an overall mean value
of 1.18. The ratio of fluoranthene to pyrene in D0, OS1, and
O8 locations (range: 0.83–0.99) were less than 1.0, whereas
the other parts (range: 1.06–2.13) had fluoranthene to pyrene
ratios of greater than 1.0. The indeno[1,2,3-cd]pyrene to ben-
zo[ghi]perylene ratios were less than 1.0 for W2, D1, D3, and
O5 locations (range: 0.12–0.69). These results suggest that the
sources of PAHs to Onsan Bay were both petrogenic and py-
rolytic. The northern locations, which are near petrochemical
industries, may receive more petrogenic inputs, whereas the
outer bay locations may be influenced by pyrolytic inputs.
Each sample was tested as both RE and fractionated extract
in the in vitro bioassays [19]. Luciferase and protein assays
were conducted after 72 h incubations. Sample responses, ex-
pressed as mean relative luminescence units over three rep-
licate wells, were converted to relative response units, ex-
pressed as a percentage of the maximum response observed
for 2,3,7,8-tetrachlorodibenzo-
p-dioxin (TCDD; %-TCDD-
max) or 17 -estradiol (E , %-E -max) standard curves gen-
␤
2
2
erated on the same day [13,18–19]. This was done to normalize
responses for day-to-day variability in response magnitude.
The mean solvent control response was subtracted from both
sample and standard responses on a plate-by-plate basis before
conversion to a percentage, to scale values from 0 to 100%-
standard-max. Significant responses were defined as those out-
side the range defined by three times the standard deviation
(
expressed in %-standard-max) of the mean solvent control
response (0%-standard-max). Total protein in the wells was
used as an index of cell number to detect outliers that were
not apparent by simple visual inspection.
Mass balance analysis (or potency balance analysis) was
used to examine whether or not the known concentration or
composition of a sample (identified by instrumental analyses)
could account for the magnitude or potency of biological re-
sponse observed [19]. Further details of in vitro bioassay and
mass balance analysis have been described elsewhere
[
13,14,19,20].

Trace organic contaminants in aqueous media
Environ. Toxicol. Chem. 21, 2002
1799
Atmospheric transportation and deposition of PAHs from the
nearby Ulsan Bay, where PAH concentrations as great as 3,100
ng/g dry weight were detected in sediments, may be a source
of PAHs to Onsan Bay [6].
Concentrations of PCBs in sediments were generally less
than 10.0 ng/g dry weight (Table 1). Locations D3 and D4 in
Daejeong Stream contained relatively great concentrations of
PCBs (19.6 and 56.2 ng/g dry wt, respectively). The spatial
gradient of PCB concentrations in sediments suggests the pres-
ence of sources along Daejeong Stream. Lesser chlorinated
congeners such as tetra- and penta-CBs were the most prev-
alent homologs in Onsan Bay sediments. Previous studies have
also reported the presence of lower chlorinated PCB congeners
in sediments collected from Masan and Ulsan bays [5,6].
Among different OC pesticides analyzed, DDT concentrations
(
sum of p,p
phenyldichloroethylene, and -DDT) were the greatest, ranging
from 0.01 to 7.58 ng/g dry weight. Concentrations of target
Ј-dichlorodiphenyldichloroethane, -dichlorodi-
Ͻ
organic compounds in the streams were greater than those in
the bay (Table 1). The likely sources of these persistent organic
pollutants are from industrial activities or municipal waste-
waters discharged from Onsan City.
Nonylphenol, OP, and BPA were detected at some locations
in Onsan Bay (Table 1). However, concentrations of these
contaminants were generally close to the method detection
limit (1.00 ng/g dry wt). Maximum concentrations of NP and
OP in sediments were 860 and 11 ng/g dry weight, respec-
tively, at location D1 in Daejeong Stream. The greater con-
centrations of APs at locations D1, D3, and D4 can be ex-
plained by its proximity to sewage waste input near D1 lo-
cation. The BPA was found in sediment from the inland river
and stream only and ranged from
Ͻ1.00 to 204 ng/g dry
weight. Similar to that for APs, the greatest concentration of
BPA was measured in sediment from location D1.
Comparison to other studies
Several studies have examined the occurrence and distri-
bution of persistent organic pollutants such as PCBs, OC pes-
ticides, and PAHs in Korean coastal areas [5–11,18]. The PCB
concentrations in sediment samples from Onsan Bay (n ϭ 3)
collected in 1995 ranged from 4.5 to 8.2 ng/g dry weight [21].
Sedimentary PCBs and PAHs in Korea have been reported to
range from a few nanograms per gram to several micrograms
per gram dry weight (Table 2). Concentrations of PCBs and
PAHs in Onsan Bay were approximately 1.5- to 10-fold less
than those reported in other industrialized coastal areas such
as Kyeonggi, Masan, Ulsan, and Yeongil bays. Concentrations
of APs and BPA in sediment from Onsan Bay were similar to
those in Ulsan Bay but less than those in other bay areas (Table
2
). Greater concentrations of APs and BPA in Lake Shihwa
and Masan and Yeoingil bays are consistent with greater input
from heavily populated cities such as Shiheung, Masan, and
Pohang. The data presented here establish the baseline for
future monitoring of these compounds in Onsan Bay areas.
Potential for biological and ecological effects
Sixteen priority PAH compounds including aryl hydrocar-
bon receptor (AhR)–active compounds such as ben-
zo[
zo[
a
]anthracene, chrysene, benzo[
b]fluoranthene, ben-
k
]fluoranthene, benzo[ ]pyrene, indeno[1,2,3-cd]pyrene,
a
and dibenz[a,h]anthracene were detected in sediments. Non-
ortho-coplanar PCB congeners 77 (3,3 ,4,4 -tetraCB), 126
3,3 ,4,4 ,5-pentaCB), and 169 (3,3 ,4,4 ,5,5 -hexaCB) also
Ј
Ј
(
Ј
Ј
Ј
Ј
Ј

1
800
Environ. Toxicol. Chem. 21, 2002
C.-H. Koh et al.
Table 3. Instrumentally derived 2,3,7,8-tetrachlorodibenzo-
p
-dioxin (TCDD) equivalents (TEQs) and predicted and observed percentage of the
maximum response for TCDD (%-TCDD-max) of Onsan Bay sediments, Korea
TEQs (pg/g dry wt)a
%-TCDD-max-calculated
PAH
%-TCDD-max-observedb
Location TEQPCB
TEQPAH
TEQs
PCB
24.1
Total
24.2
F1
F2
RE
W0
W1
W2
D0
D1
D2
D3
D4
D6
ON1
ON2
ON3
OS1
OS2
OS3
O4
O5
O6
O7
O8
3.276
0.019
0.016
8.859
NDc
6.113
0.003
3.249
0.009
ND
0.009
0.340
4.928
ND
0.414
ND
3.285
0.359
4.944
8.859
0.414
6.113
0.154
4.601
0.252
ND
Ͻ
Ͻ
0.00
0.00
0.47
2.43
9.76
0.60
0.00
3.59
8.54
22.8
48.7
85.0
29.1
14.2
7.8
17.1
16.9
Ͻ
Ͻ
0.00
0.00
Ͻ
0.00
31.9
42.6
0.00
27.2
0.00
29.5
0.00
31.9
NAC
42.6
NAC
27.2
d
Ͻ
0.00
Ͻ
Ͻ
2.31
4.14
23.6
18.7
32.7
2.61
NA
NA
Ͻ
0.00
0.00
0.00
0.00
NAC
Ͻ
Ͻ
Ͻ
21.5
3.06
12.5
9.1
0.151
1.352
0.242
ND
Ͻ
0.00
23.2
0.00
NAC
Ͻ
0.00
7.30
0.00
Ͻ
14.5
Ͻ
Ͻ
Ͻ
5.54
0.00
NA
NA
2.49
NA
NAC
NA
NA
NAC
NA
NA
e
0.348
ND
NA
0.348
NA
Ͻ
0.00
NAC
NAC
NAC
NAC
NA
ND
ND
ND
0.260
0.461
0.159
0.252
0.434
0.150
0.003
ND
0.260
0.461
0.159
0.289
0.463
0.150
0.003
ND
Ͻ
0.00
0.00
0.00
0.00
0.00
0.00
0.00
Ͻ
0.00
0.00
0.00
0.00
0.00
0.00
0.00
15.9
NA
NA
38.0
44.9
28.4
NA
NA
NA
22.6
14.6
20.9
18.3
20.1
36.6
15.7
11.1
12.7
14.7
18.1
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
NA
0.037
0.029
ND
Ͻ
Ͻ
0.00
0.00
2.61
3.41
1.51
NA
NA
NA
NAC
NAC
NAC
NAC
NAC
ND
ND
ND
ND
NAC
NAC
O9
O10
0.028
0.170
0.028
0.170
Ͻ
Ͻ
0.00
0.00
Ͻ
Ͻ
0.00
0.00
0.60
a
Instrumentally derived TEQs of polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) associated with sediment
samples. Refer to the TEQs generated from assay-specific median relative potency factor values and selected PCB concentrations (PCBs 81,
7
7, 118, 105, 126, 167
ϩ
185, 156, and 169) and PAH concentrations (benzo[
a
]anthracene, chrysene, benzo[
b]fluoranthene, benzo[k]fluoranthene,
benzo[ ]pyrene, indeno[1,2,3-cd]pyrene, and dibenz[a,h]anthracene).
a
b
c
d
e
F1
ϭ
least polar fraction (contains PCBs); F2
ϭ mid-polar fraction (contains PAHs); RE ϭ raw extract.
ND
ϭ not detected.
NAC
NA
ϭ
not available for calculation.
ϭ
not analyzed.
were detected in a few samples. Based on the concentrations
of dioxinlike compounds, 2,3,7,8-TCDD equivalents (TEQs)
were estimated by using relative potency factors specific to
the H4IIE-luc assay for selected PAHs and non- and mono-
ortho-PCB congeners [19]. Concentrations of total TEQs
ranged from less than the method detection limit to 8.86 pg/
g dry weight (Table 3). The TEQPCBs for 50% of the locations
were not detected, whereas TEQPAHs for all the locations except
for four (D0, D2, ON1, and O8) were detected. Contributions
of PCBs and PAH concentrations to total TEQs varied among
locations. This result suggested that both dioxinlike PCBs and
PAHs were responsible for the H4IIE-luc responses observed
in sediment RE samples. The TEQs estimated for selected PCB
and PAHs in Onsan sediment were at the lower end of the
sediment quality guideline of 0.014 to 210 pg/g dry weight
reported for dioxin equivalents [22].
Sediment quality guidelines such as effect range low (ERL)
and threshold, median, and extreme effects concentrations for
PAHs and PCBs were applied to further evaluate the quality
of sediment in Onsan Bay [23–25]. The total organic carbon–
normalized concentrations of 16 individual PAHs, total PAHs,
and total PCBs were calculated. None of the locations ex-
ceeded the ERLs for 16 PAHs and PCBs. When threshold
effects concentrations reported for total PCBs were compared,
only one location (D4) exceeded the limit of 35 ng/g dry
weight. Because the ERL determined for PAHs and PCBs is
influenced by co-occurring toxicants in field studies, it may
be an over estimate of the toxicity attributable to PAHs or
PCBs. This results in an artificially low value for the ERL.
Thus, calculation of hazard quotients based on ERLs would
overestimate the effects. In any case, PAH or PCB concentra-
tions did not exceed the ERLs, thus neither PAHs nor PCBs
would be expected to have adverse effects on benthic inver-
tebrates.
Dioxinlike activity in vitro
Extracts of sediment, water (dissolved fraction and partic-
ulate matter), and pore water were screened for their ability
to induce AhR-mediated gene expression in vitro with H4IIE-
luc cells [19]. Based on the initial screening of raw extracts,
17 of 22 sediment samples showed significant dioxinlike ac-
tivity in the H4IIE-luc bioassay (Table 4). Surface water and
pore-water samples were less active. No pore-water samples
produced a significant response in the H4IIE-luc bioassay.
Among the surface water samples, only samples from locations
D2 and D5 elicited a significant response. Response magni-
tudes ranged from 11.0%-TCDD-max to 17.5%-TCDD-max
and responses for the particulate matter did not differ signif-
icantly from those from the dissolved fraction sample. The
activity of the D2 and D5 water samples could not be explained
based on the instrumental data available. The unusual bioassay
responses suggest that these areas may warrant further study
as potential locations containing point sources or relatively
hydrophilic AhR agonists. Overall, the RE screening results
suggested that most dioxinlike organic contaminants in the
Onsan Bay area aquatic environment were associated with
sediment. This is consistent with the relatively hydrophobic
properties of most known AhR agonists.
To examine potential cause–effect relationships between the
target (known) AhR agonists quantified in this study and the
AhR-mediated bioassay responses observed, sediment extracts
were divided into three fractions and each fraction was ana-

Trace organic contaminants in aqueous media
Environ. Toxicol. Chem. 21, 2002
1801
a
Table 4. Luciferase bioassay response (%-TCDD-max) of H4IIE-luc cells for the sediment raw extracts (RE) and fractionated samples (F1, F2,
and F3) from Onsan Bay, Korea
Location
RE
F1b
F2c
F3d
W0
W1
W2
D0
D1
D2
D3
D4
D6
ON1
ON2
ON3
OS1
OS2
OS3
O4
14.22
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
4.38
4.49
1.04
3.28
0.40
0.72
3.21
4.99
2.61
1.20
3.72
6.86
5.45
4.92
1.98
1.98
4.09
1.98
4.88
2.79
3.79
0.72
0.47
2.43
9.76
0.60
0.00
3.59
8.54
14.52
5.54
0.00
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
2.63
2.02
2.90
4.91
4.73
3.10
5.11
2.67
2.22
13.02
24.56
29.28
4.61
25.22
4.80
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
0.63
5.33
5.55
2.92
1.19
0.76
3.86
2.15
6.14
3.23
60.32
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
5.97
1.85
6.70
8.83
0.32
0.69
6.06
15.98
3.29
1.45
7.78
17.10
16.87
13.91
54.09
52.14
Ͻ
0.00
0.00
0.00
0.00
Ͻ
Ͻ
Ͻ
0.00
0.00
Ͻ
Ͻ
Ͻ
6.76
15.55
23.07
42.12
27.47
1.39
21.47
3.06
12.51
9.09
Ͻ
Ͻ
0.00
0.00
Ϯ
2.49
Ϯ
e
NA
NA
Ϯ
NA
NA
Ϯ
Ϯ
Ϯ
NA
NA
NA
Ϯ
NA
NA
Ϯ
NA
NA
Ϯ
Ϯ
Ϯ
NA
NA
NA
Ϯ
NA
NA
14.60
20.90
18.28
20.11
36.64
15.65
11.12
12.73
14.68
18.13
2.79
1.13
5.31
16.39
16.47
Ϯ
NA
NA
3.15
2.61
3.41
1.51
1.99
2.33
2.73
7.60
6.25
9.89
9.38
7.46
6.61
9.21
12.14
8.41
Ϯ
Ϯ
Ϯ
0.87
1.70
5.93
O5
O6
O7
O8
NA
NA
NA
O9
O10
Significant level
0.60
2.60
4.05
2.30
12.87
Ϯ
4.36
f
4.56
2.87
2.87
2.87
a
Response magnitude presented as percentage of the maximum response observed for a 2,000 pM 2,3,7,8-tetrachlorodibenzo-
standard.
p-dioxin (TCDD)
b
c
d
e
f
F1
F2
F3
NA
ϭ
ϭ
ϭ
fraction 1 of sediment samples (contains PCBs).
fraction 2 of sediment samples (contains PAHs).
fraction 3 of sediment samples (contains alkylphenols and bisphenol A).
not analyzed.
ϭ
Significant level is defined as 99% confidence interval around the mean solvent control response (set to 0%-TCDD-max).
lyzed in the H4IIE-luc assay. Previously reported spike-re-
covery tests [5] have shown AhR-active PCBs to partition to
the least polar fraction (F1) and AhR-active PAHs to partition
to the mid-polar fraction (F2). High-resolution mass spectrom-
D6, and O5) that should not have yielded a F1 response did
yield a response, and two samples (W0 and D0) that should
have yielded F1 responses did not do so. Furthermore, the
maximum magnitude of response observed was 14.5%-TCDD-
max (Table 3), which was significantly lower than the mag-
nitude expected for all four predicted responses. Thus, re-
eter analysis suggests that polychlorinated dibenzo-p-dioxins
and polychlorinated dibenzofurans partition primarily to F2,
with only a small percentage occurring in F3 [13]. Thus, if
the target, AhR-active analytes for this study were the pre-
dominant causes of AhR-mediated reporter gene expression in
vitro, one would expect F1 and F2 to have the greatest activity,
whereas that of F3 should be minimal. Furthermore, the mag-
nitude of activity expected in F1 and F2 could be predicted
sponses of F1 did not conform well to the additive, TEQPCB
,
model applied. This suggests that either non–AhR-active PCB
components of F1 were modulating the activity of the AhR-
active PCBs, or that the AhR-active PCBs present were acting
in a nonadditive fashion in the H4IIE-luc bioassay.
If target analytes were the primary cause of the AhR-me-
diated bioassay responses observed for Onsan area sediment
samples, TEQPAH should have resulted in activity in F2. The
additive TEQPAH model predicted that only two F2 samples
(W2 and D4) contained sufficient concentrations of dioxinlike
PAHs to induce a significant response in the H4IIE-luc bio-
assay (Table 3). Contrary to this prediction, 13 of the 15 F2
samples analyzed yielded a significant response (Table 3).
Twelve of the 14 responses were in excess of 20%-TCDD-
max (Table 3). The F2 samples for both W2 and D4 yielded
significant responses, as predicted, but the response magni-
tudes observed were approximately 2.6-fold greater than pre-
dicted (Table 3). The strong and consistent deviation from the
additive TEQPAH model suggests that PAHs were not the pri-
mary cause of activity in F2 or that AhR-active PAHs were
acting in a greater than additive manner. Because polychlor-
(
Table 3) based on the measured concentrations of AhR-active
PCBs, AhR-active PAHs, and their previously reported, H4IIE-
luc-specific, relative potencies [19,26]. Predicted magnitudes
of response were based on the assumption that TEQs would
behave in the bioassay as if they were pure 2,3,7,8-TCDD
standard and that all TEQs acted in a simple additive fashion
(
no synergistic or antagonistic interactions). Predicted and ob-
served magnitudes of response were compared in a qualitative
mass (potency) balance to evaluate whether or not the target
analytes quantified in this study could account for the respons-
es observed.
Based on TEQPCB concentrations present in the samples, 4
of the 15 F1 samples analyzed should have yielded a significant
H4IIE-luc response (Table 3). Each of those significant re-
sponses was predicted to be 20%-TCDD-max or greater (Table
3
). However, this predicted profile of responses was not ob-
inated dibenzo-p-dioxins and polychlorinated dibenzofurans
served. Instead, 6 of the 15 F1 samples analyzed produced a
significant response (Table 4). Only two of these (D2 and D4)
were samples predicted to yield a significant F1 response,
based on TEQPCB concentrations. Thus, four samples (W2, D3,
are known to partition predominantly to F2, one likely expla-
nation for the greater than predicted responses is that AhR-
active dioxins and furans were present in the Onsan area sed-
iment samples. Polychlorinated naphthalenes are another class

1
802
Environ. Toxicol. Chem. 21, 2002
C.-H. Koh et al.
a
Table 5. Luciferase bioassay response (%-E -max) of MCF-7-luc (MVLN) cells for the sediment raw extracts (RE) and fractionated samples
2
(
F1, F2, and F3) from Onsan Bay, Korea
F1b
Location
RE
0.00
F2c
F3d
W0
W1
W2
D0
D1
D2
D3
D4
D6
ON1
ON2
ON3
OS1
OS2
OS3
O4
Ͻ
Ͻ
Ͻ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
11.2
10.7
2.37
9.84
2.15
18.9
9.19
4.42
2.31
22.8
4.80
4.62
1.51
1.87
4.74
6.17
3.54
4.15
6.58
2.80
5.85
5.55
1.62
5.50
8.86
4.75
0.00
3.86
0.50
2.65
1.48
0.83
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
2.63
2.02
2.90
4.91
4.73
3.10
5.11
2.67
2.22
13.0
24.6
29.3
4.61
25.2
4.80
6.76
15.5
0.00
0.00
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
1.11
3.93
4.65
1.83
4.98
4.21
1.01
3.32
0.92
1.54
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
0.00
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
Ϯ
1.19
0.80
1.44
2.41
0.69
4.28
4.33
1.34
5.37
0.32
0.00
0.00
0.15
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
Ͻ
Ͻ
Ͻ
10.44
Ͻ
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ͻ
Ϯ
2.49
Ϯ
e
NA
NA
Ϯ
NA
NA
Ϯ
Ϯ
Ϯ
NA
NA
NA
Ϯ
NA
NA
Ϯ
NA
NA
Ϯ
Ϯ
Ϯ
NA
NA
NA
Ϯ
NA
NA
3.44
1.13
5.31
2.88
Ͻ
Ͻ
0.00
Ϯ
NA
NA
2.45
3.44
5.03
8.77
1.99
2.33
2.73
7.60
6.25
9.89
3.31
0.53
6.11
1.67
0.00
1.99
Ϯ
Ϯ
Ϯ
2.07
2.60
1.17
O5
O6
O7
O8
NA
NA
NA
0.001
0.00
0.00
O9
O10
Significant level
Ͻ
Ͻ
1.62
2.60
4.05
1.46
Ͻ
0.00
Ϯ
1.09
f
6.60
1.32
1.32
1.32
a
Response magnitude presented as percentage of the maximum response observed for a 1,000 pM 17
␤
-estradiol (E ) standard.
2
b
c
d
e
f
F1
F2
F3
NA
ϭ
ϭ
ϭ
fraction 1 of sediment samples (contains PCBs).
fraction 2 of sediment samples (contains PAHs).
fraction 3 of sediment samples (contains alkylphenols and bisphenol A).
not analyzed.
ϭ
Significant level is defined as 99% confidence interval around the mean solvent control response (set to 0%-E -max).
2
of ubiquitous, dioxinlike compounds that may partition to F2
relative potencies do not account for the type of interactions
implied by these results. Additional identification and char-
acterization of novel AhR agonists and antagonists may im-
prove future evaluations.
[
15] and may have contributed to the F2 responses. Such com-
pounds should be investigated in future studies of persistent
organic pollutants in the Onsan area.
No known AhR-agonists were expected to partition into F3.
Despite this, 12 of the 15 F3 samples analyzed produced a
significant response in the H4IIE-luc bioassay (Table 4). The
prevalence and magnitude of AhR activity in F3 suggests the
presence of unidentified, relatively polar, AhR agonists in sed-
iment from the Onsan area. The results are consistent with
previous studies, which examined sediments from other Ko-
rean coastal areas [13,18,19]. Thus, the unidentified agonists
seem to have a fairly ubiquitous distribution near Korea. Future
studies should employ extensive bioassay-directed fraction-
ation and chemical analysis in an effort to identify the caus-
ative agents in F3 samples from this study and other locations
in Korea. Additional research also could be aimed at deter-
mining whether similar F3 responses would be observed for
sediments from other areas, both contaminated and relatively
uncontaminated, worldwide.
Estrogenic activity in vitro
Onsan area sediment extracts also were screened for their
ability to induce estrogen receptor–mediated gene expression
in vitro [19]. Only one RE sample (D3) yielded a significant
response in the MVLN bioassay (Table 5). The general lack
of response was consistent with the relatively low concentra-
tions of known estrogen receptor agonists in extracts of Onsan
area sediment (Table 1). Based on MVLN-specific relative
potencies previously determined for NP, OP, BPA, and two
weakly estrogenic PAHs (benzo[a]anthracene and di-
benz[a,h]anthracene), concentrations of E equivalents based
2
on these residues in the extracts were not predicted to elicit a
response in the MVLN bioassay.
Analysis of fractionated extracts complicated the picture
somewhat. Twelve out of 15 F1 samples and 13 of 15 F2
samples caused significant estrogenic responses (Table 5).
Magnitudes of MVLN response for F2 samples as great as
Comparison of RE and fractionated extract sample respons-
es suggested that a simple additive model considering pres-
ently characterized AhR agonists is probably not sufficient to
characterize the AhR-mediated toxic potency of the complex
mixtures of compounds found in Korean sediment. Among the
29.3%-E -max (W2) were observed (Table 4). This is equiv-
2
alent to a response elicited by 21 pM of E . Based on the PAH
analysis in sediment samples, concentrations of estrogenic
2
1
5 samples tested as both RE and fractionated extract, 4 of
PAHs such as benzo[a]anthracene (65.3 ng/g dry wt) and di-
F1, 14 of F2, and 9 of F3 samples yielded fractionated extract
responses that were greater than the corresponding RE re-
sponses, respectively (Table 4). This implies antagonistic in-
teractions within the total extract that were relieved by sep-
arating components of the complex mixture. Current additive
models for predicting dioxinlike potency based on congener-
specific chemical concentrations and previously determined
benz[a,h]anthracene (8.73 ng/g dry wt) in W2 were greatest
among samples analyzed. The E equivalent for estrogenic
2
PAHs in sediment from W2 was 4.01 pM of E based on the
2
reported PAH concentrations and their relative potency values
[26]. This mass balance suggested that concentrations of es-
trogenic PAHs account only for a portion of estrogenic re-
sponses observed in F2. Thus, the F2 results suggest that Onsan

Trace organic contaminants in aqueous media
Environ. Toxicol. Chem. 21, 2002
1803
by polycyclic aromatic hydrocarbons in Ulsan Bay (South Korea)
sediments. Environ Pollut 111:437–445.
. Chang YS, Kong SB, Ikonomou MG. 1999. PCBs contributions
to the total TEQ released from Korean municipal and industrial
waste incinerators. Chemosphere 39:2629–2640.
9. Kim SK, Lee DS, Oh JR, Kahng SH. 2000. Effects of extreme
tidal range on characteristics of polychlorinated biphenyl distri-
bution in sediment of industrial Incheon North Harbor, Korea.
Environ Toxicol Chem 19:2448–2456.
10. Lee KT, Tanabe S, Koh CH. 2001. Contamination of polychlor-
inated biphenyls (PCBs) in sediments from Kyeonggi Bay and
nearby areas, Korea. Mar Pollut Bull 42:273–279.
area sediments, from at least some locations, contain estrogen
receptor–active compounds that elute in the same Florisil frac-
tion as do PAHs. Additional bioassay-directed fractionation
and chemical analysis would be necessary to identify the caus-
ative agents. No F3 samples yielded a significant MVLN re-
sponse. Instead, W1, D1, D3, and D4 F3 samples, all river or
stream sediments, were cytotoxic to the MVLN cells. The F3
results suggest a significant difference in the chemical profile
of inland river and stream sediment and coastal sea sediment.
Overall, in vitro bioassay applied in conjunction with instru-
mental analysis provides a powerful tool for characterizing
mechanism-specific agonists present in the environment.
8
1
1. Lee KT, Tanabe S, Koh CH. 2001. Distribution of organochlorine
pesticides in sediments from Kyeonggi Bay and nearby areas,
Korea. Environ Pollut 114:207–213.
1
1
2. Nimrod AC, Benson WH. 1996. Environmental estrogenic effects
of alkylphenol ethoxylates. Crit Rev Toxicol 26:335–364.
3. Khim JS, Villeneuve DL, Kannan K, Koh CH, Giesy JP. 1999.
Characterization and distribution of trace organic contaminants
in sediment from Masan Bay, Korea: 2. In vitro gene expression
assays. Environ Sci Technol 33:4206–4211.
4. Hilscherova K, Machala M, Kannan K, Blankenship AL, Giesy
JP. 2000. Cell bioassays for detection of aryl hydrocarbon receptor
(AhR) and estrogen receptor (ER) mediated activity in environ-
mental samples. Environ Sci Pollut Res 7:159–171.
5. Kannan K, Villeneuve DL, Yamashita N, Imagawa T, Hashimoto
S, Miyazaki A, Giesy JP. 2000. Vertical profiles of dioxin-like
and estrogenic activities associated with a sediment core from
Tokyo Bay, Japan. Environ Sci Technol 34:3560–3567.
SUMMARY
The relative abundance of organic contaminants measured
in Onsan Bay was in the order PAHs
ഠ APs Ͼ BPA Ͼ PCBs
Ͼ
OC pesticides. Spatial distributions of these contaminants
1
1
in Onsan Bay suggested that their sources were independent
of each other. Overall, the target organic contaminants ex-
amined in this study do not seem to be a severe problem in
Onsan Bay. Bioassay responses to F1 in the H4IIE-luc cell
bioassay were consistent with the relatively low concentrations
of dioxinlike PCBs detected in Onsan Bay sediments. Mass
balance analysis suggested that PAHs accounted for only a
portion of dioxinlike activity in F2 samples. The undefined
components of F2 and F3 seem to be responsible for the ma-
jority of the dioxinlike responses. The lack of responses of
MVLN cells to samples was consistent with the low concen-
trations of estrogenic compounds such as NP, OP, and BPA in
Onsan Bay sediments. Based on mass balance analysis, the
known estrogenic PAH and AP composition of F3 probably
accounts for less than 20% of the estrogenic activity observed
in the MVLN bioassay. Overall, these results suggest the need
for more extensive fractionation and instrumental analysis to
identify uncharacterized AhR and estrogen receptor agonists
and cytotoxic compounds in F3 samples.
16. Carr RS, Chapman DC. 1995. Comparison of method for con-
ducting marine and estuarine sediment porewater toxicity tests—
Extraction, storage, and handling techniques. Arch Environ Con-
tam Toxicol 28:69–77.
1
1
1
7. Khim JS, Villeneuve DL, Kannan K, Hu WY, Giesy JP, Kang
SG, Song KJ, Koh CH. 2000. Instrumental and bioanalytical mea-
sures of persistent organochlorines in blue mussel (Mytilus edulis
from Korean coastal waters. Arch Environ Contam Toxicol 39:
60–368.
)
3
8. Khim JS, Villeneuve DL, Kannan K, Lee KT, Snyder SA, Koh
CH, Giesy JP. 1999. Alkylphenols, polycyclic aromatic hydro-
carbons (PAHs), and organochlorines in sediment from Lake
Shihwa, Korea: Instrumental and bioanalytical characterization.
Environ Toxicol Chem 18:2424–2432.
9. Khim JS, Lee KT, Villeneuve DL, Kannan K, Giesy JP, Koh CH.
2001. In vitro bioassay determination of dioxin-like and estro-
genic compounds in environmental samples from Ulsan Bay and
its vicinity, Korea. Arch Environ Contam Toxicol 40:151–160.
Acknowledgement—This study was financially supported by the Na-
tional Institute of Environmental Research, Ministry of Environment,
Korea, Sediment Organic Compounds Bioassay Study 1998–2000.
20. Yamashita N, Kannan K, Imagawa T, Villeneuve DL, Hashimoto
S, Miyazaki A, Giesy JP. 1999. Vertical profile of polychlorinated
dibenzo-p-dioxins, dibenzofurans, naphthalenes, biphenyls, poly-
cyclic aromatic hydrocarbons, and alkylphenols in a sediment
core from Tokyo Bay, Japan. Environ Sci Technol 34:3560–3567.
1. Kim GW, Lee Y. 1996. Concentrations of PCBs (polychlorinated
biphenyls) in coastal sediments of Korea. Korean J Sanit 11:9–
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