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Author's personal copy
Chemosphere 69 (2007) 164–169
www.elsevier.com/locate/chemosphere
Concentrations of pentachlorophenol (PCP) in fish and shrimp
in Jiangsu Province, China
Jiachun Ge
a,b,*
,
Jianling Pan
a,c
, Zhiliang Fei
a,b
, Guanghong Wu
a,b
, John P. Giesy
d,e,f
a
f
Freshwater Fishery Research Institute of Jiangsu Province, China, 79 Chating East Street, Nanjing 210017, China
b
Fishery Analysis and Testing Center of Jiangsu Province, China
c
School of Life Science, Nanjing University, China
d
Department of Veterinary Biomedical Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan, Canada
e
National Food Safety and Toxicology Center, Department of Zoology and Institute for Environmental Toxicology,
Michigan State University, East Lansing, MI 48824, USA
Center for Coastal Pollution and Conservation, Department of Biology and Chemistry, City University of Hong Kong, HK SAR, PR China
Received 13 November 2006; received in revised form 20 March 2007; accepted 4 April 2007
Available online 29 May 2007
Abstract
Pentachlorophenol (PCP) concentrations were determined in 55 samples of fish, shrimp, crabs, frogs and turtles collected in Jiangsu
Province and 55 freshwater shrimp samples collected from the Huiming Fish Market, Nanjing, China between 2003 and 2004. PCP concentrations ranged from less than the method detection limit (MDL: 0.5 lg/kg ww) to 61 lg/kg ww, with a mean of 5.2 lg/kg ww in 55
samples collected across Jiangsu Province and a mean of 0.5 lg/kg ww for freshwater shrimp collected from the Huiming Fish Market.
Concentrations of PCP in common carp (Carassius auratus) were significantly greater than concentrations of PCP in other species from
all of the areas studied. Grass carp (Ctenopharyngodon idella) contained the second greatest concentrations and crustacean species contained the least concentrations. Concentrations of PCP were greater in samples collected from Southern Jiangsu than those collected
from other areas of Jiangsu Province. Concentrations of PCP in the freshwater shrimp (Macrobrachium nipponense) from the Huiming
Fish Market were greater in March and September than those collected in June and December. Based on the allowable daily intake
(ADI) set by the US National Academy of Science, current concentrations of PCP measured in aquatic products from Jiangsu Province
presented minimus health risks to humans.
2007 Elsevier Ltd. All rights reserved.
Keywords: Pentachlorophenol; Sodium pentachloronate; Fishery products; Jiangsu; Risk assessment
1. Introduction
Pentachlorophenol (PCP) has been used as an herbicide,
algaecide, defoliant, wood preservative, germicide, fungicide and molluskacide (Jorens and Schepens, 1993). In
1997, China produced approximately 104 tonnes of PCP
per year, which was approximately 20% of global production (Zhang et al., 1997).
PCP is readily absorbed across the skin, lungs and gastrointestinal lining (Reigart and Roberts, 1999). The Uni*
Corresponding author. Tel.: +86 25 86581570; fax: +86 25 86618250.
E-mail address: gjc09@hotmail.com (J. Ge).
0045-6535/$ - see front matter 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.chemosphere.2007.04.025
ted States Environmental Protection Agency (USEPA)
has determined that PCP is a probable human carcinogen
but the International Agency for Cancer Research (IARC)
considers it to be only a possible carcinogen in humans
(Agency for Toxic Substances and Disease Registry USA,
2001). Currently the regulatory status of PCP varies among
countries. PCP is registered in the United States of America
(USA) as a ‘‘restricted use’’ pesticide for use only as a wood
preservative (Jorens and Schepens, 1993). Use of PCP
ceased in Austria, India, Indonesia, New Zealand, Sweden,
Switzerland or Germany at different times between 1978
and 1991 and use has been restricted in the European
Union and Belize. In China, PCP is only allowed as a wood
Author's personal copy
J. Ge et al. / Chemosphere 69 (2007) 164–169
preservative (FAO, 1996). Since the 1960s, PCP was used
as a molluskacide in China, but this application was
stopped in 2000 (Tao, 2003).
From the 1960s until it was banned, PCP was used
extensively in Jiangsu Province, as the major molluskacide
to kill Oncomelania hupensis which is the intermediate host
of schistosomiasis (Wang et al., 1998). In order to investigate the possible risk of PCP in freshwater fishery products
of humans of Jiangsu Province, China through dietary
exposure, concentrations of PCP were measured in 110
samples of freshwater fish, shrimp, crabs, frogs and turtles
collected from across Jiangsu Province and measured concentrations were compared to allowable daily intake (ADI)
values.
2. Methods
2.1. Sample collection
In July, September and October 2003 and January 2004,
55 samples of shrimp (Macrobrachium nipponense, Macrobrachium rosenbergii and Penaeus Vanmamei), Chinese mitten crab (Eriocheir sinensis), fish including common carp
(Carassius auratus), big head carp (Aristichthys nobilis),
silver carp (Hypophthalmichthys molitrix), grass carp (Ctenopharyngodon idella), bull frog (Rana catesbeiana) and
turtle (Amyda sinensis) were collected in aquaculture farms
or markets in the cities of Suzhou, Changzhou, Wuxi, Nanjing, Huaian, Lianyungang and Yangzhou in Jiangsu Province. In June, September and December 2003 and March
2004, 11, 15, 15 and 14 samples, respectively, of the shrimp
(M. nipponense) were also collected at the Huiming Fish
Market, Nanjing, Jiangsu, China. The muscle of shrimp
abdomen, fish, bull frog and turtle, and the liver, muscle
and gonad of crab were taken, homogenized, and used
for determination of PCP concentrations.
2.2. Quantification of PCP
PCP was quantified by use of extraction into organic
solvent, followed by quantification by gas chromatography
with electron capture detection (ECD). Samples (1 g ww)
were homogenized with sulfuric acid (8 ml; 50%) in a
homogenizer (T-18, IKA, Guangzhou, China), then kept
at 80 C in a water bath for 30 min. The homogenate was
extracted 2 times with n-hexane (10 ml each). The n-hexane
phase was extracted 2 times with potassium carbonate solution (0.1 M, 4 ml each). Acetic anhydride (0.2 ml) was
added to the potassium carbonate phase, and mixed.
The mixture was then extracted 2 times with n-hexane
(2 + 2.5 ml). The n-hexane was removed to a 5 ml pyknoflask and the volume adjusted to 5 ml before analysis by
use of gas chromatography with electron capture detector.
Solvent blanks and QA samples were included with each
batch of samples. Recoveries of PCP from spiked samples
were tested on a reference shrimp sample (M. nipponense)
from the Yangtze River and a reference cod fish sample
165
from a supermarket in Nanjing. Average recoveries were
77–92% from when the shrimp was spiked with 1.0–
5.0 lg PCP/kg ww and 78–94.8% from the cod fish spiked
with 2.0–10.0 lg PCP/kg ww. A standard solution of PCP
was purchased from the China National Research Center
for Certified Reference Materials. The solvent used in the
studies, n-hexane, was HPLC or ‘‘Spectro’’ grade (Tedia
Company USA), sulfuric acid was G.R., potassium carbonate, acetic anhydride and sodium sulfate (dried at
500 C for 5 h) were all analytical grade.
Concentrations of PCP in extracts were determined by
use of an Agilent 6890 gas chromatograph equipped with
63
Ni micro-electron capture detector based on the methods
of Fei et al. (2004) and Ge et al. (2006). The column was a
DB-35MS 30 m · 0.25 mm · 0.25 lm. Injection was splitless and injector and detector temperatures were 250 C
and 300 C, respectively. The temperature program was
140 C for 2 min, 10 C min 1 to 200 C isothermal for
7 min, then 230 C for 3 min. The method detection limit
(MDL) was defined as a value 3 times as great as the blank
signal.
3. Results and discussion
Concentrations of PCP in the 55 samples from Jiangsu
Province ranged from less than the MDL (0.5 lg PCP/
kg ww) to 61 lg PCP/kg, (common carp from the Wuxi
area). The mean PCP concentration was 5.2 lg PCP/
kg ww. Concentrations of PCP were less than 1.0 lg
PCP/kg ww for 54.5% of the samples, while 36.4% of samples contained concentrations between 1.0 lg PCP/kg ww
and 10 lg PCP/kg ww. Only 5 samples (9%) contained
PCP concentrations greater than 10 lg PCP/kg ww. The
samples containing the greatest concentrations of PCP
included one grass carp and four common carp. Among
the four common carp concentrations were greater than
50 lg PCP/kg ww for three of them (Table 1). Ranges of
mean concentrations of PCP in the various species are
given along with the area from which the samples were collected and the proportion of samples in the range (Table 2).
Concentrations of PCP in M. nipponense from the
Huiming Fish Market, Nanjing, Jiangsu, collected in
different seasons, ranged from less than the MDL to
1.4 lg PCP/kg ww in 55 shrimp. Concentrations of PCP
were greatest in September 2003, with a mean concentration of (0.7 ± 0.3 lg PCP/kg ww), followed by samples
collected in March 2004 (0.6 ± 0.3 lg PCP/kg ww), then
by samples collected in December 2003 (0.3 ± 0.2 lg
PCP/kg ww). Samples collected in June 2003 contained the
least concentrations of PCP (0.25 ± 0.00 lg PCP/kg ww)
(Fig. 1). There were no statistically significant differences
between samples collected in September or March or
between samples from December or June (Duncan’s test,
a = 0.05). However, concentrations of PCP in samples
collected during September or March were significantly
greater than those of samples collected in December and
June (Duncan’s test, a = 0.01).
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166
J. Ge et al. / Chemosphere 69 (2007) 164–169
Table 1
Ranges of PCP concentrations and proportions in the range 55 samples from Jiangsu Province, China
PCP concentration range
(lg PCP/kg ww)
Number of
samples
Sample name
Percentage
<0.5
0.5–1.0
1.0–10
23
7
20
41.8
12.7
36.4
10–50
50–60
>60
2
2
1
Chinese mitten crab, M. nipponense, grass carp, P. Vanmamei, M. rosenbergii
Chinese mitten crab, M. nipponense, grass carp, big head carp
Chinese mitten crab, M. nipponense, grass carp, M. rosenbergii, silver carp,
big head carp, bull frog, turtle
Grass carp, common carp
Common carp
Common carp
Table 2
Mean PCP concentration of species, by area of collection
Wuxi
Samples
Chinese mitten crab
Grass carp
Common carp
n
3
3
3
Average
Changzhou
Chinese mitten crab
M. nipponense
Common carp
Silver carp
Big head carp
Bull frog
Turtle
4
3
1
1
4
1
1
1.5
PCP average
concentration
(lg/kg)
SD
2.0
6.9
52
0.68
5.2
8.0
20
25
1.2
1.0
55
2.4
2.4
2.5
2.5
1.2
1.3
–
–
2.6
–
–
Mean PCP Concentration (of ug PCP /Kg, ww)
Area
3.6
3.6
1.8
1.4
1.3
1.2
1.1
1.0
.9
.8
.7
.6
.5
.4
.3
.2
.1
0.0
June 2003
Sep. 2003
Dec. 2003
March 2004
Season
Average
Suzhou
Chinese mitten crab
M. rosenbergii
M. nipponense
3
3
3
Average
Nanjing
Chinese mitten crab
Grass carp
3
3
Average
5.3
14
1.2
1.8
2.0
0.50
0.11
0.20
1.7
0.50
0.42
0.37
0.18
0.17
0.40
0.16
Nantong
M. nipponense
2
0.29
0.06
Huaian
M. nipponense
1
0.56
–
Taizhou
M. nipponense
4
0.27
0.05
Yangzhou
M. rosenbergii
3
0.25
0.00
Lianyungang
P. Vanmamei
6
0.25
0.00
3.1. Comparison of PCP concentrations among species
Common carp contained the greatest PCP concentration
(53 ± 6.6 lg PCP/kg ww, n = 4), followed by grass carp
(3.6 ± 4.9 lg PCP/kg ww, n = 6), big head carp (2.4 ±
2.6 lg PCP/kg ww, n = 4), Chinese mitten crab (1.2 ±
0.9 lg PCP/kg ww, n = 13), M. rosenbergii (1.0 ± 0.9 lg
PCP/kg ww, n = 6), M. nipponense (0.9 ± 0.9 lg PCP/
kg ww, n = 13) and P. Vanmamei (0.25 ± 0.00 lg PCP/
Fig. 1. PCP concentration of shrimp M. nipponense samples from
Huiming Fish Market, Nanjing in different seasons.
kg ww, n = 6). Concentrations of PCP in common carp
were significantly greater than those of all other species
(Duncan’s test, a = 0.01). No statistically significant differences in PCP concentrations were observed among grass
carp, big head carp, Chinese mitten crab, M. rosenbergii
and M. nipponense. Concentrations of PCP in grass carp
were significantly greater than those of P. Vanmamei
(Fig. 2). Because the number of samples of some species
was limited, concentrations of PCP were reported for
these species, they were not included in the statistical
comparisons.
Concentrations of PCP were also compared among species within the following locations: Changzhou, Wuxi and
Suzhou, because the greatest numbers of samples were collected from these locations (Table 2). In the Changzhou
area, PCP concentrations were greatest in common carp
(55 lg PCP/kg ww, n = 1), followed by big head carp
(2.4 ± 2.6 lg PCP/kg ww, n = 4), Chinese mitten crab (1.2 ±
1.2 lg PCP/kg ww, n = 4) and M. nipponense (1.0 ±
1.3 lg PCP/kg ww, n = 3). There were no statistically significant differences among big head carp, Chinese mitten
crab and M. nipponense. The mean PCP concentration in
common carp (52 ± 8.0 lg PCP/kg ww, n = 3) from Wuxi,
was significantly greater than that of the grass carp
(6.9 ± 5.2 lg PCP/kg ww, n = 3) and Chinese mitten crab
Author's personal copy
Mean PCP Concentration (of ug PCP/Kg, ww)
J. Ge et al. / Chemosphere 69 (2007) 164–169
concentration of PCP in crustaceans was 0.9 ± 0.8 lg PCP/
kg ww. The average concentration of PCP in fish was
16 ± 23 lg PCP/kg ww. Concentrations of PCP were significantly greater in fish than in crustaceans (T-test, a =
0.05). The reason could be the differences in trophic level
and possibly differences of lipid content or metabolism.
In two crustacean species Gammarus pulex and Daphnia
magna the bioaccumulation factors (BCF) for PCP are 51
(Ashauer et al., 2006) and 35 (Tan and Sun, 1994), respectively, less than that in 3 fish species Oryzias latipes, Pimephales promelas and Lepomis macrochirus (Tachikawa et al.,
1991; Huckins and Petty, 1983; Pruitt et al., 1977) (Table 3).
It seems that some fish species can accumulate PCP faster
than some crustacean species. More information is not
available yet.
70
60
50
40
30
20
10
0
-10
P.
se
ei
am
nm
Va
n
ne
po
ip
.n
en
itt
m
rp
ca
ii
rg
be
en
os
.r
M
M
ad
he
rp
ca
rp
ca
se
ne
hi
C
g
Bi
ss
ra
G
on
m
om
C
ab
cr
Species
167
3.2. Comparison of PCP concentrations among areas
Fig. 2. Differences of PCP concentration among samples across Jiangsu.
(2.0 ± 0.7 lg PCP/kg, n = 3) (Duncan’s test, a = 0.01). The
mean concentration of PCP in grass carp was greater than
that of the Chinese mitten crab, but the difference was not
statistically significantly (Duncan’s test, a = 0.05). In the
Suzhou area, average PCP concentrations of three species
of crustacean were relatively less than those of other locations, all of them were less than 2.0 lg PCP/kg ww. Among
these species, the mean PCP concentration of M. nipponense (2.0 ± 0.2 lg PCP/kg, n = 3) was significantly greater
than that of the Chinese mitten crab (1.2 ± 0.5 lg PCP/
kg ww, n = 3), but the difference between M. nipponense
and M. rosenbergii (1.8 ± 0.1 lg PCP/kg ww, n = 3) was
not significant (Duncan’s test, a = 0.05). Similarly, there
was no statistically significant difference in PCP concentration between M. rosenbergii and Chinese mitten crab. Fish
contained greater concentrations of PCP than did crustaceans collected from the same area.
Concentrations of PCP were also compared among
crustaceans (M. nipponense, M. rosenbergii, Chinese mitten
crab and P. Vanmamei) and fish (common carp, big head
carp and grass carp) across Jiangsu Province. The average
PCP concentrations in samples from Wuxi (n = 9),
Changzhou (n = 15), Suzhou (n = 9), Nanjing (n = 6),
Nantong (n = 2), Taizhou (n = 4), Yangzhou (n = 3) and
Lianyungang (n = 6) were compared. The samples from
Wuxi contained the greatest concentrations of PCP with
a mean concentration of 20 lg PCP/kg ww. The average
PCP concentration in samples from Wuxi was significantly
greater than that of samples from Suzhou, Nanjing, Nantong, Taizhou, Yangzhou or Lianyungang, but the difference between Wuxi and Changzhou was not statistically
significant (Duncan’s test, a = 0.05) (Fig. 3).
Because the above comparisons involved all species, and
the species occurred in different areas and at different proportions, it was difficult to make definitive conclusions
about the trends among areas. The Chinese mitten crab,
M. nipponense and grass carp were chosen to compare the
difference among areas. Concentrations of PCP in the Chinese mitten crab were statistically greater in samples from
Wuxi than those from Nanjing, followed by Suzhou,
Changzhou and Nanjing (Duncan’s test, a = 0.05). As for
M. nipponense, the greatest mean concentration of PCP
was found in Suzhou, followed by Changzhou, Nantong,
Taizhou, while that at Suzhou was significantly greater than
Table 3
PCP uptake and elimination data of some fish and crustacean species
Species
Uptake rate constant
(ml g 1 h 1)
Depuration rate
constant (h 1)
Bioaccumulation factors
Reference
Gammarus pulex
Daphnia magna
Medaka (Oryzias latipes)
3.7 ± 0.3
0.073 ± 0.006
51
35
20.8–10.6
0.0097–0.0170
Ashauer et al. (2006)
Tan and Sun (1994)
Tachikawa and
Sawamura (1994)
Tachikawa et al.
(1991)
Huckins and Petty
(1983)
Pruitt et al. (1977)
Medaka (Oryzias latipes)
Fathead minnows
(Pimephales promelas)
Bluegill sunfish (Lepomis
macrochirus)
Hyalella azteca
Freshwater and seawater acclimated fish:
1680 and 370, respectively
For purified PCP: 174, for industrial
composite: 284
10–350
25.7 ± 2.9
0.061 ± 0.034
Nuutinen et al. (2003)
Author's personal copy
Mean PCP Concentration (of ug PCP/Kg, ww)
168
J. Ge et al. / Chemosphere 69 (2007) 164–169
70
60
50
40
30
20
10
0
Li
Ya
an
u
ng
u
ga
o
zh
u
n
yu
ng
ho
iz
g
ng
jin
to
an
o
zh
u
ng
an
Ta
N
N
ha
i
ux
o
zh
Su
C
W
Sampling Area
Fig. 3. Comparison of PCP concentration of samples among different
areas in Jiangsu.
that of Nantong and Taizhou (Duncan’s test, a = 0.05). For
the grass carp, for which only two areas could be compared,
the concentration of PCP in samples from Wuxi were significantly greater than in samples from Nanjing (T-test, one
side, a = 0.05). Concentrations of PCP in samples of aquatic
organisms from South Jiangsu (Wuxi, Suzhou, Changzhou)
were slightly greater than those of samples from other areas
of Jiangsu Province. The primary reason for this is that PCPNa was used widely as a molluskacide in South Jiangsu
Province to control schistosomiasis by killing O. hupensis.
The total amount of PCP applied in Jingtan county of
Changzhou of South Jiangsu between 1970 and 1993 was
64 tonnes, which represents an application rate of approximately 10 g PCP/m2, was greater than the amount applied
in other areas (Wang et al., 1998).
There is little information in the literature about the
concentrations of PCP in aquatic animals in China. In
2000, PCP residues were found to range from less than
the MDL to 0.63 mg PCP/kg in freshwater fish bile in
Dongting Lake in China (Zheng et al., 2000). Concentrations of PCP in freshwater sediments and human breast
milk collected from the Pearl River Delta, China have been
reported (Hong et al., 2005). The concentration of PCP in
river sediments ranged from 1.4 to 34 ng PCP/g with a
mean of 7.9 ng PCP/g, dry weight. The average concentration of PCP in human breast milk was 2.2 ng PCP/g ww,
and was directly proportional to age of donors.
3.3. Potential risk of PCP in food to humans
The allowable daily intake (ADI) value for PCP has
been established by the US National Academy of Science to
be 3 lg PCP/kg bw/day (WHO, 1989). Until now, Codex/
JMPR (2006) has not established an ADI or maximum residue limit (MRL) for PCP in food consumed by humans. In
several countries an MRL in the human diet has been
established for PCP. For instance the MRL set by Belgium
for PCP in mushrooms is 50 lg PCP/kg ww. In Germany
for all foods of plant origin, it is 10 lg PCP/kg ww. In
Israel for mushrooms and other foods, it is 50 lg PCP/
kg ww. In Netherlands, it is 50 lg PCP/kg ww for mushrooms, while in Switzerland, it is 50 lg PCP/kg ww for
milk and in Yugoslavia for unnamed commodities, it is
10 lg PCP/kg (FAO, 1996). The strictest MRL set by those
countries for any commodity is 10 lg PCP/kg.
In the survey conducted in Jiangsu Province only 9% of
the 55 samples from across the province exceeded
10 lg PCP/kg ww, while none of the 55 samples of shrimp
from the Huiming Fish Market, in Nanjing exceeded this
value. Assuming the body weight of an average adult to be
60 kg and that each person consumes, on average, 0.25 kg
fish and shellfish/day, and that the PCP concentration in
fish and shellfish is as great as the greatest concentration observed in this survey (mean = 53 lg PCP/kg ww
in common carp), the PCP exposure of humans would
be 0.22 lg PCP/kg bw/day, which is approximately 7% of
the ADI (3 lg PCP/kg bw/day) suggested by the US
National Academy of Science. Thus, based on either the
MRL or ADI, the current level of exceedence for the human
population in Jiangsu Province would be less than 10%. This
suggests that the level of risk to humans is relatively small
and no immediate actions need to be taken to restrict exposure of the public.
Acknowledgments
This study was financially supported by Jiangsu Science
and Technology Department (BS2002031 and BG2005324), China. The authors wish to thank Wu Jun, Li Ping,
Zhang Meiqing, Shen Meifang and Gao Yuyi, who helped
conduct the residue analyses.
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