Monitoring of Endocrine Disruptor Compounds in the
Coastal Hydrosphere of
R. Boonyatumanond, A. Jaksakul, S.Boonchalermkit,
P.Puncharoen,
M.S. Tabucanon
Environmental Research and
Technopolis, Tambon Klong 5, Amphoe Klong Luang,
Department of Environmental Quality Promotion,
Ministry of Science, Technology and
Environment
ABSTRACT
The
contamination of Endocrine Disruptor Compounds (EDCs) in the environment has been recognized as the important issue of
pollution problem. These compounds are
occurred from industrial products or
industrial wastes which have been known as PCBs, dioxins, pesticides including
organochlorine, phenols, phthalates, etc.
In order to identify the situation of EDCs contamination, the Environmental Research and Training
Center has cooperated with The United Nations University (UNU) to establish the
monitoring program of EDCs in the coastal hydrosphere of Thailand during
1999-2002 under the UNU project on " Environmental and Governance:
Endocrine Disruptor Compounds Pollution in the East Asian Coastal
Hydrosphere. In 2001, fourty eight
samples of water were collected from rivers and sea along the coastal area in the upper
INTRODUCTION
Many Endocrine Disrupting Chemical (EDCs) are regarded as
organic pollutants in the environment which occurred from industrial chemicals,
pesticides and by-products of manufacturing processes as well as products of
incineration of industrial and household wastes. The known EDCs include polychlorinated
biphenyl (PCBs), organochlorine pesticides (OCPs) and other types of
pesticides, dioxins, alkylphenol polyethoxylates and organotins compounds.
These chemicals are discharged into the environment through industrial
wastewater, municipal waste, agricultural runoffs and eventually flow into
rivers and coastal areas. Many kind of
EDCs such as diethyl phthalate (DEP) and di-butyl phthalate (DBP) which used as
a plasticizer, solvent for resins, wetting agent and insect repellent may
release into the environment through wastewater effluent. Bis 2-ethylhexyl
phthalate(DEHP) use as an insulating fluid in electrical transformers ,
industrial tubing and food packaging system including bis2-ethylhexyl adipate
which released into the environment during PVC blending operations and consumer
used of finished product. DBP is also exposed from using cosmetics and food
wrapping . Phenol is widely used in the manufacturing process such as
resins, plastic, insecticide, explosives, dye and detergent, etc.
Organochlorine pesticides have been extensively used in agricultural
activities. In
EDCs have been found in
contaminated food, in polluted drinking water and in some plastics which
tainted with EDCs.Human is exposed to
EDCs through oral ingestion, contamination of the skin or inhalation through
the lungs. Many EDCs are able to
accumulate in fatty tissue of living organisms due to their lipophilic
characteristic. EDCs may affect the
normal functions of
endocrine system on
reproduction , cancers and immunological system.
According
to the effects of EDCs on human, there is a need to conduct monitoring and
analysis of the contamination of these chemicals in the environment . The Environmental Research and Training
Center has cooperated with The United Nation University (UNU) to establish the
monitoring program of EDCs in the coastal hydrosphere of Thailand during
1999-2002 under the UNU project on " Environmental and Governance :
Endocrine Disruptor Compounds Pollution in the East Asian Coastal Hydrosphere.
The objective is to identify the situation of EDCs contamination in water along
the coastal area and the main
MATERIALS AND METHODS
The monitoring programme started from April-November
2001. Fourty eight samples of water were collected in both wet and dry seasons
at 24 stations from four main rivers namely, the Chao Phraya river, the
Mae-Klong river, the Bang Pa-kong river and the Tha-Chin river including the
coastal areas along the gulf of Thailand and Andaman sea side as showned in Figure 1, Figure 2
and Table
1. The samples were analysed for
organochlorine pesticide, phenol compounds and phathalate compounds by
following UNU method. All water samples
were kept at 4 °C and especially for phenol
analysis the samples were preserved with HCL to pH 2-3.
APPARATUS
A)
Gas-Chromatograph-model
17-A (Shimadzu) equipped with Mass Spectrometer model QP-5000 (Shimadzu )
B)
Rotary Vacuum evaporator-type Rotavapor RE-111
(Buchi)
C)
Laboratory sharking machine-type Recipo shaker
SR-IIW
(Taitec
corperation)
D)
Chromatographic column-capillary 30m x 0.32 mm
id x 0.25 mm of film thickness DB5 ( J&W
)
E)
Vortex-type model
GENIE 2 G-560E (Scientific Inc.,
F)
Glassware-Separatory
funnel 2000ml with glass stopper; glass funnel 90 mm id; cylinder graduated
50ml; Erlenmeyer flask (Pyrex)
REAGENTS
a)
Solvent-Ultra
high pure grade (J.T. Baker Chemical Inc.U.S.A.)
b)
Silica-gel
cartridge 500mg (Varian)
c)
Organochlorine
pesticides standard ( AccuStandard Inc. and Wako chemical industrial Ltd. )
-
a-HCH
-
b-HCH
-
g-HCH
-
d-HCH
-
p,p’-DDE
-
p,p’-DDT
-
p,p'-DDD
-
Aldrin
-
Dieldrin
-
Endrin
10 mg of each standard was weighted accurately on an
analytical balance, put it in volumetric flask with 100ml and dissolved with
100ml n-hexane-acetone for stock solution.
d)
Internal standard
solution (
-
Phenanthrene d10
-
Pyrene d10
-
Naphthalene d8
-
Di n-butyl
phthalate d4
-
Di 2-ethyl hexyl
phthalate d4
e)
Phenol standard
(Wako chemical industrial Ltd.,)
-
4-t-Butylphenol
-
2,4-Dichlorophenol
-
4-n-Butylphenol
-
4-n-Pentylphenol
-
4-n-Hexylphenol
-
4-n-Heptylphenol
-
4-t-Octylphenol
-
4-n-nonyltylphenol
-
4-n-Octylphenol
-
Pentachlorophenol
-
Bisphenol-A
-
Bisphenol-A d14
f)
sodium sulfate (
g)
sodium chloride (
h)
Phthalate
standard
-
Di-ethyl
phthalate
-
Di n-butyl
phthalate
-
Di 2-ethyl hexyl
adipate
-
Di 2-ethyl hexyl
phthalate
Sample preparation for organochlorine pesticides
1 liter of water sample added
with 30 g of sodium chloride, was extracted with 50 ml of n-hexane for 10
minutes by shaker. The hexane was
transferred to Erlenmeyer flask. The water sample was extracted repeatedly with
50ml of n-hexane. After shaking, the
extractant was transferred into the same flask.
The hexane was dehydrated with sodium sulfate until the volume was reduced
into 1ml. and then transferred to
silica gel cartridge for cleanup. The
cartridge was washed with 5 ml of acetone and 15ml of n-hexane. Organochlorine pesticide residues was eluted
with 5ml of 5% acetone/n-hexane and was injected into GC/MS for detection. This
method and QA/QC were followed the UNU manual 1999.
Sample preparation for phenol compounds
500ml of water sample was adjusted pH condition to 2-3
by using hydrochloric acid and add 30 g sodium chloride into separatory funnel.
The sample was added with100 ml of 1ppm of surrogate compound (Bisphenol-A d16) and was extracted with
25 ml of dichloromethane by shaker for 10 min
. The organic layer was
transferred to erlenmeyer flask and the water was repeatedly extracted again
with 25 ml dichloromethane. The organic
layer was transferred into the same flask. The extractant was dehydrated with
sodium sulfate and concentrated to 0.5 ml by rotary evaporator and
nitrogen. The extracting was added 100 ml BSTFA and stay for 1 hour at room temperature and
added 100 ml of internal standard
before inject into GC/MS.
Sample preparation for phthalate compounds
Add 100ml of water sample
into a 100ml volumetric flask and add 5
ml of n-hexane. The internal standards
were added 10ml of 10ppm and shake for 1 min for
extraction. Transfer 1 ml of hexane
layer from volumetric flask into vial for injection into GC/MS
GC-MS confirmation
Mass
fragmentation data were obtained with Gas Chromatograph model GC-17A (Shimadzu)
and interface equipped with Mass Spectrometer model QP-5000 (Shimadzu) and
Electron impact mode (ion energy was 70 EV).
The chromatography column bonded DB-5 fuse silica column (30m x 0.32mm
id x 0.25mm film thickness) was used.
The condition set for
organochlorine pesticide compounds
as follows :
The injection method was
splitless 2 min, injection volume was 2 ml and injection inlet temperature was 280 °C. The carrier gas was helium with a flow rate of 2
ml/min. The initial column temperature
was 70°C(2min) and increase at
20°C/min to 150°C, increase at 5 °C/min to 220 °C and increase at 15 °C/min to 300 °C. interface temperature at 280°C
The condition set for phenol compounds as follows :
The injection method was
splitless 2 min, injection volume was 2 ml and injection inlet temperature was 300 °C. The initial
column temperature was 50 °C (2 min) and increase at 20°C/min to 200°C, increase at 20°C/min to 300°C (8 min)., interface temperature at 270°C
The condition set for
phthalate compounds as follows :
The injection method was
splitless 2 min, injection volume was 2 ml and injection inlet temperature was 320 °C. The initial
column temperature was 70 °C (2 min) and increase at 10°C/min to 200°C, and increase 5°C to 245° C/min and 20°C to 320°C (3 mim),. interface temperature at 300°C
RESULTS AND DISCUSSIONS
Forty-eight water samples were
analyzed for 10 organochlorine pesticides by following UNU method.
The method detection limit are
3.0-17 ng l-1(ppt)
with recovery of
78-88%. In Table 2 5 the results were shown that 8
from 48 samples , the organochlorine pesticides
residues were detected of b-HCH (nd-10 ppt), d-HCH (nd-14 ppt), dieldrin (nd-25 ppt), p,p'-DDE
(nd-15 ppt), p,p'-DDD (nd-17 ppt), and p,p'-DDT( 20 ppt). The highest
concentration of organochlorine pesticides residue in river water samples were
detected of dieldrin and p,p'-DDD at the Tha-Chin river
in dry season (TR1, 25 ppt) and the Chao phraya river in wet season(CH5, 3.0
ppt), respectively. The highest concentrations of organochlorine pesticides
residue in sea water samples were detected p,p'-DDT, p,p'-DDD at Krabi( 20 ppt)
and Samutprakarn(17 ppt), respectively. b-HCH, d-HCH and p,p'-DDE were detected only 1 from 48 samples at Pattani, Prachub-kirikorn and Krabi
station, respectively.
The
river water and sea water samples were monitored of 11 phenol compounds in dry
season and wet season. Phenol residues were found 4-t-butylphenol,
4-n-pentylphenol, 4-n-butylphenol, 4-nonylphenol, 2,4-dichlorophenol and
Bisphenol A. The results were shown in Table 6 - 9. The method detection limit
are 0.01-0.06 ng ml-1(ppb) and 76-111% recovery. The detected samples of 4-t-butylphenol were about 66% of
total sample. The concentration of 4-t-butylphenol were at
the range of nd-2.5 ppb. The highest residue of river water sample and
sea water sample were detected in dry season at the Tha-Chin river and Nakhorn srithamarat , respectively. The
concentration of 4-n-pentylphenol, and 4-n--butylphenol were detected at the range of nd-0.02 ppb and detected only one
station at Krabi in wet and dry season ( 0.02 ppb). The detected samples of 4-n-pentylphenol and 4-n--butylphenol were about 0.04% of total sample. The concentration range of 2,4
dichlorophenol was nd-0.02 ppb, The detected
samples of 2,4-dichlorophenol were about 25% of total sample, especially
in river water sample. The concentration
range of 4-nonylphenol was nd-0.85 ppb and the highest concentration was 0.85
ppb at The Tha-Chin river station in wet season . The concentration of Bisphenol-A was
detected at the range of nd-0.32 ng ml-1 . The
highest concentration was found 0.32 ppb at The Tha-Chin river in dry season.
The detected samples of Bisphenol-A
detection was about 89% of total sample.
The result of phenol
concentration in water sample is very low concentration because behaviour of
phenol such as high vapor pressure.
Those compounds can photooxidize
in the air and about 90% degradation in surface water. Although the data showed
that many water samples found phenol residues but the concentration is lower
than the limit of the water quality
standard in Thailand which is limit <0.005 mg l-1 or 5 ng ml-1.
In
case of Phathalate residues analysis, they were selected 4 compounds by UNU
method , di ethyl phthalate (DEP), di n-butyl phthalate (DBP), di 2-ethylhexyl
phthalate (DEHP) and di 2-ethylhexyl adipate (DEHA). The results were shown in
Table 10 - 13. It indicated
that only DEHP and DEHA were detected . The detected samples of DEHA and DEHP were 0.04% and 14% of total samples
respectively. The concentration range of DEHA was nd-0.04 ppb which was
detected only one station at the Tha-Chin river station in dry season . The concentration range of DEHP was nd-0.09
ppb. The highest concentration was detected
0.09 ppb at the Tha-Chin river station in dry season . DEP could not be
detected , it might because of the chemical property such as slowly volatilize
from water surface and can adsorb into sediment or particulate matter .
Conclusion
During April
2001 March 2002, The
Environmental Research and Training
Center of Thailand has
cooperation with the United Nations
University to monitor organochlorine pesticides , phenols
and phthalates in freshwater and
seawater .Eleven stations of four main rivers
which flow to the Upper Gulf of Thailand ;namely Chao Praya , Mae Klong
, Tha Chin , Bang Pakong ; and thirteen stations of coastal areas have been
selected for monitoring and analysis. Fourty
eight samples were collected in wet and dry season..
The results of organochlorine
pesticides monitoring indicated that most of the samples were not detected
except in some sample were detected of b-HCH, d-HCH dieldrin, p,p'-DDE , p,p'-DDD p,p'-DDT at the concentration range of 10 20 ngl -1
(ppt) which are lower than the water quality standard.
The results
of phenol monitoring indicated
that the samples collected from river
and seawater of 24 stations were detected of
4 nonylphenol at the
concentration range of 0.04 2.1 ngml 1 (ppb) . About 66% of total samples were detected of 4 t Butylphenol at the concentration
range of
0.02 2.5 ppb. About 89% of total samples were detected of Bisphenol A at the concentrations range
of 0.02 0.32 ppb. About 27 %
of total samples were detected of
2 , 4 Dichlorophenol at
the concentrations range of
0.01 0.02 ppb including 4 n
Butylphenol and 4 n
Pentylplenol were detected in 2 samples of seawater
at the concentration of 0.02 ppb. It was shown that the concentration of phenol was lower than
the water quality standard which is
limit at 5 ngml 1
The results
of phthalates monitoring
along rivers and coastal areas
indicated that 8 from 48 samples were detected of di 2 ethylhexyl phthalate ( DEHP) at the concentration range from 0.007 0.09 ngml 1 (ppb) while only 1 sample
was detected of di 2 ethylhexyl adipate ( DEHA ) at the
concentration of 0.04 ppb.
Figure 1 Map of sampling stations (coastal area)
Figure 2 Map of sampling stations (River Basin)
Table 1 Location
of sampling stations
|
Number of sample |
Code number |
Name of station |
Remark |
|
1 |
CH1 |
The Chao Praya river |
-1 : dry season |
|
2 |
CH2 |
The Chao Praya river |
-2 : wet season |
|
3 |
CH3 |
The Chao Praya river |
|
|
4 |
CH4 |
The Chao Praya river |
|
|
5 |
CH5 |
The Chao Praya river |
|
|
6 |
MK1 |
The Mae Klong river |
|
|
7 |
MK2 |
The Mae Klong river |
|
|
8 |
TR1 |
The Tha Chin river |
|
|
9 |
TR2 |
The Tha Chin river |
|
|
10 |
BP1 |
The Bang pakong river |
|
|
11 |
BP2 |
The Bang pakong river |
|
|
12 |
TRAD |
Trad province |
|
|
13 |
CHONB |
Chonburi province |
|
|
14 |
SMP |
Samutprakarn province |
|
|
15 |
PATTANI |