Page 1
Enantiomeric Excess of POPs in the Environment
Hiramoto S1, Tsurukawa M2, Matsumura C2, Nakano T2, Kunugi M3
1Hyogo Environmental Advancement Association2Hyogo Prefectural Institute of Public Health Association3National Institute for Environmental Studies
Page 2
IntroductionIntroduction
Page 3
Purpose
To identify the behavior of POPs
Enantiomeric compositions of chiral POPs in seawater and air were investigated.
The results of chiral analysis were shown using Enantiomeric Excess (EE).
* In this study, (+) or (-) enantiomer does not identi
fied.
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Enantiomeric Excess(EE)
Changes in physicochemical status,
It is possible to distinguish between newly caused pollution and preserved by monitoring EE
Changes in metabolic status,
the EE value does not change.
the EE value changes.
Metabolites showed enantioselective degradation of (+) or (-) enantiomer.
Page 5
Materialsand
Methods
Materialsand
Methods
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S-1
S-2
S-3
S-4
S-5
S-6
S-7
S-8
S-9
S-10
S-11
May~ June 2006
Sampling Sites (Seawater)~ South China Sea ~
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P-1
P-2
P-3
P-4
P-5
P-6
P-7
Sampling Sites (Seawater)~ Pacific Ocean (West) ~
August~ September 2005
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P-8
P-9
P-10P-11
August~ September 2005
UNITED STATES
San Francisco
Sampling Sites (Seawater)~ Pacific Ocean (East) ~
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N-1
N-2
N-3
N-4
May~ June 2005
Sampling Sites (Air)~ North Atlantic Ocean ~
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Marine pollution observation systemMarine pollution observation system
Solid PhaseExtraction unit
Control Unit
ExtractionColumn
Filter unit
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Sample Preparation for Analysis
Exchange to hexaneExchange to hexane
Clean up with silica gelClean up with silica gel
Sample ( PUF +ACF )
Sample ( PUF +ACF )
Soxhlet Extraction 3hr with acetone 24hr for dichloromethane
Soxhlet Extraction 3hr with acetone 24hr for dichloromethane
Concentration
Concentration , Dehydration
HRGC/HRMSHRGC/HRMS
Concentration
Polyurethane foam
( PUF )
Glass holder
Active carbonfiber filter( ACF )
Spiked with 13C-labeld POPsSpiked with
13C-labeld POPs
Air 400m3 Seawater 100L
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Target Analytes
trans - Chlordane
Heptachlor α - HCH
Heptachlor epoxideOxychlordane
o,p’ - DDT
o,p’ - DDE
cis - Chlordane
Technical compounds
o,p’ - DDD
Metabolites
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HRGC/HRMSGC MS
Device : HP-6890N(HP)
Column : BGB-172*(BGB Analytik)
30m, 0.25mm i.d. film thickness 0.25μm
Carrier Gas : He 1.5mL/min
Device : JMS-800D (JEOL)
Ionization : EI
Ion source temperature :260 ℃
Resolution : >10,000Temperature Program : 120 (2min) → 2 /min →250 (3min)℃ ℃ ℃Injector temperature : 230℃ Injection Volume : 2μL
* BGB-172 ( 20% tert-butyldimethylsilyl-β-cyclodextrin dissolved in 15% diphenyl-polysiloxane and 85% dimethylpolysiloxane )
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HP-5
8 9 10 11 12 13 14 15Retention Time (min)
(3448523)
0
25
50
75
100
Inte
nsity
HCH / Average
α
β γ δ
36 38 40 42 44 46 48 50Retention Time (min)
(1082345)
0
25
50
75
100
Inte
nsity
HCH / Average
α
βγ δ
BGB-172
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Enantiomeric ExcessEnantiomeric Excess (EE)
EE(%) = *100
Enantiomeric Excess (EE)
EE(%) = *100
EL-ES
EL+ES
EL : amount of larger enantiomerES : amount of smaller enantiomer
0
20
40
60(%)
EE
(+)
(-)
(+)
(-)
(+)(-)
(+)
(-)
(+)
(-)EE= 0 %
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33 34 35 36 37 38 39 40 41 42 43 44 45 46 47Retention Time (min)
(81468)
0
25
50
75
100
Inte
nsi
ty
P18-13 (Unknown sample)33 34 35 36 37 38 39 40 41 42 43 44 45 46 47
Retention Time (min)
(2053941)
0
25
50
75
100
Inte
nsi
ty
STDmix-4 (Calibration standard)
35 36 37 38 39 40 41 42 43 44 45 46 47 48 49Retention Time (min)
0
200000
Inte
nsi
ty
POPs2 ATLAS-22 (Unknown sample)
35 36 37 38 39 40 41 42 43 44 45 46 47 48 49Retention Time (min)
0
40000
80000
Inte
nsi
ty
POPs2 Air-6 (Unknown sample)
HCHalpha-beta-
gamma-
delta-
EE = 1%
EE = 6%
EE = 10%
EE = 3%
standard
Seawater ( South China Sea)
Seawater ( Pacific Ocean)
Air (North Atlantic Ocean)
RACEMIC
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57.8 58.0 58.2 58.4 58.6 58.8 59.0 59.2 59.4 59.6 59.8 60.0 60.2 60.4 60.6
Retention Time (min)
4000
8000
Inte
nsi
ty
POPs2 Air-8 (Unknown sample)
55.0 55.2 55.4 55.6 55.8 56.0 56.2 56.4 56.6 56.8 57.0 57.2 57.4 57.6 57.8 58.0Retention Time (min)
(2054268)
0
25
50
75
100
Inte
nsi
ty
STDmix-4 (Calibration standard)
Chlordanetrans-
cis-
EE = 1%
EE = 1%
trans- EE = 4%
cis- EE = 7%
standard
Air (North Atlantic Ocean)
54.8 55.0 55.2 55.4 55.6 55.8 56.0 56.2 56.4 56.6 56.8 57.0 57.2 57.4 57.6Retention Time (min)
(1003)
(14412)
0
25
50
75
100
Inte
nsity
chlordane / Average
57.6 57.8 58.0 58.2 58.4 58.6 58.8 59.0 59.2 59.4 59.6 59.8 60.0 60.2 60.4Retention Time (min)
(529)
(4236)
0
25
50
75
100
Inte
nsity
chlordane / Averagetrans- EE = 4%
cis- EE = 1%
Seawater ( Pacific Ocean)
RACEMIC
trans- EE = 4%
cis- EE = 5%
Seawater ( South China Sea)
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52.8 53.2 53.6 54.0 54.4 54.8 55.2 55.6 56.0 56.4 56.8 57.2Retention Time (min)
(1095846)
0
25
50
75
100
Inte
nsi
ty
STDmix-4 (Calibration standard)
51.6 52.0 52.4 52.8 53.2 53.6 54.0 54.4 54.8 55.2 55.6 56.0Retention Time (min)
(25007)
0
25
50
75
100
Inte
nsi
ty
P18-16 (Unknown sample)
55.6 56.0 56.4 56.8 57.2 57.6 58.0 58.4 58.8 59.2 59.6 60.0Retention Time (min)
0
2000
4000
Inte
nsi
ty
POPs2 Air-8 (Unknown sample)55.2 55.6 56.0 56.4 56.8 57.2 57.6 58.0 58.4 58.8 59.2 59.6 60.0
Retention Time (min)
1000
2000
3000
Inte
nsi
ty
POPs2 ATLAS-22 (Unknown sample)
Heptachlor epoxideendo-
exo-
EE = 1%
EE = 1%
EE = 39%
EE = 28%
EE = 44%
standard
Seawater ( South China Sea)
Seawater ( Pacific Ocean)
Air (North Atlantic Ocean)
NON
RACEMIC
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53 54 55 56 57 58 59 60 61 62Retention Time (min)
(6369865)
0
25
50
75
100
Inte
nsi
ty
STDmix-4 (Calibration standard)
53 54 55 56 57 58 59 60 61 62Retention Time (min)
(75117)
0
25
50
75
100
Inte
nsi
ty
P18-13 (Unknown sample)
55 56 57 58 59 60 61 62 63 64Retention Time (min)
0
20000
Inte
nsi
ty
POPs2 ATLAS-10 (Unknown sample)
55 56 57 58 59 60 61 62 63 64Retention Time (min)
4000
8000
Inte
nsi
ty
POPs2 Air-8 (Unknown sample)
DDDo,p’- p,p’-
EE= 1 %
EE = 51%
EE = 58%
standard
Seawater ( South China Sea)
Seawater ( Pacific Ocean)
Air (North Atlantic Ocean)
NON
RACEMIC
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52.0 52.4 52.8 53.2 53.6 54.0 54.4 54.8 55.2 55.6 56.0 56.4 56.8Retention Time (min)
(155320)
0
25
50
75
100
Inte
nsi
ty
STDmix-4 (Calibration standard)
52.0 52.4 52.8 53.2 53.6 54.0 54.4 54.8 55.2 55.6 56.0 56.4 56.8Retention Time (min)
500
1000
1500
2000
Inte
nsi
ty
POPs2 ATLAS-22 (Unknown sample)
54.8 55.2 55.6 56.0 56.4 56.8 57.2 57.6 58.0 58.4 58.8 59.2 59.6Retention Time (min)
800
1200
1600
Inte
nsi
ty
POPs2 Air-1 (Unknown sample)
52.0 52.4 52.8 53.2 53.6 54.0 54.4 54.8 55.2 55.6 56.0 56.4 56.8Retention Time (min)
(650)
(1678)
0255075
100
Inte
nsi
ty
P18-13 (Unknown sample)
OxychlordaneEE=1%
EE=1%
standard
Seawater ( South China Sea)
Seawater ( Pacific Ocean)
Air (North Atlantic Ocean)
RACEMIC
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Seawater (South China Sea)
0
20
40
60
80
o,p'-DDD heptachlorexo-epoxide
alpha-HCH trans-chlordane
cis-chlordane
EE(%)
Metabolite Technical compound
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Seawater (Pacific Ocean)
0
20
40
60
80
o,p'-DDD heptachlorexo-epoxide
alpha-HCH trans-chlordane
cis-chlordane
EE(%)
Metabolite Technical compound
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Air (North Atlantic Ocean)
0
25
50
heptachlorexo-epoxide
oxychlordane alpha-HCH trans-chlordane
cis-chlordane
EE(%)
Metabolite Technical compound
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Conclusion(1)
EE of chiral POPs in seawater and air samples have been investigated.
The metabolites; heptachlor exo-epoxide and o,p’-DDD exist non racemic.
Enatiomeric composition of alpha-HCH, trans-,cis-chlordane were near racemic.
However, oxychlordane is the metabolite, it exists as racemic.
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Conclusion(2)
By monitoring enantiomeric composition, better understanding for the mechanism of environmental pollution will be provided.
Further studies about global scale observations of chiral signatures are necessary.
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EE values (South China Sea)
S-1 S-2 S-3 S-4 S-5 S-6 S-7 S-8 S-9 S-10 S-11 averagealpha-HCH 3.8 15 5.9 7.6 6.6 1.3 7.0 2.5 5.9 8.9 5.0 6.3o,p'-DDD - 59 64 34 64 50 47 44 51 34 51 50 heptachlor exo-epoxide - - - - - - - - - - 39 39 heptachlor endo-epoxide - - - - - - - - - - - -trans - chlordane 3.3 4.1 3.3 5.0 6.1 3.8 4.2 4.3 4.2 9.0 0.9 4.4cis - chlordane 7.2 3.8 0.5 4.8 4.4 5.1 0.9 14 4.6 4.1 7.2 5.1oxychlordane - - - - - - - - - - - -
Seawater from South China Sea
0
10
20
30
40
50
60
70
S-1 S-2 S-3 S-4 S-5 S-6 S-7 S-8 S-9 S-10 S-11
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S-1S-2
S-3
S-4S-5
S-6S-7
S-8S-9
S-10
S-11
EE values (South China Sea)
0
10
20
30
40
50
60
70
S- 1 S- 2 S- 3 S- 4 S- 5 S- 6 S- 7 S- 8 S- 9 S- 10 S- 11
alpha- HCH o,p'- DDD heptachlor exo- epoxide trans- chlordane cis- chlordane
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EE values (Pacific Ocean)
P-1 P-2 P-3 P-4 P-5 P-6 P-7 P-8 P-9 P-10 P-11 averagealpha-HCH 5.4 9.0 4.3 13 13 5.4 2.2 10 10 6.3 4.3 7.6o,p'-DDD 21 59 70 64 62 58 - - - - - 56 heptachlor exo-epoxide 25 29 6.5 - 9.9 30 21 20 28 20 24 21 heptachlor endo-epoxide - - - - - - - - - - - -trans - chlordane 0.6 4.8 4.8 38 3.0 2.6 3.3 9.0 3.9 4.1 8.7 7.6cis - chlordane 7.3 6.8 1.6 9.9 8.7 7.7 4.9 19 1.0 1.5 3.3 6.5oxychlordane - - - - - - - - - - - -
Seawater from Pacific Ocean
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P-1
P-2
P-3
P-4
P-5 P-7
P-6
P-8
P-9
P-10
P-11
EE values (Pacific Ocean)
0
10
20
30
40
50
60
70
P-1 P-2 P-3 P-4 P-5 P-6 P-7 P-8 P-9 P-10 P-11
alpha-HCH o,p'-DDD heptachlor exo-epoxide trans-chlordane cis-chlordane
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EE values (North Atlantic Ocean)
N-1 N-2 N-3 N-4 averagealpha-HCH 0.7 0.1 4.5 3.0 2.1o,p'-DDD - - - - -heptachlor exo-epoxide 34 7.6 40 44 31 heptachlor endo-epoxide - - - - -trans - chlordane 4.7 16 0.4 3.8 6.2cis - chlordane 2.0 2.2 2.2 7.1 3.4oxychlordane 1.4 - 5.7 - 3.5
Air from North Atlantic Ocean
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N-1
N-2
N-3
N-4
EE values (North Atlantic Ocean)
0
10
20
30
40
50
N-1 N-2 N-3 N-4
alpha-HCH heptachlor exo-epoxide trans-chlordane cis-chlordane oxychlordane
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Persistent Organic Pollutants (POPs)
① Toxic ② Persistent in the environment ③ Bioaccumulative through the food web ④ Long-range transportable
① Toxic ② Persistent in the environment ③ Bioaccumulative through the food web ④ Long-range transportable
Stockholm Convention (May 2001)Take measures to eliminateReduce the release into the environment
Stockholm Convention (May 2001)Take measures to eliminateReduce the release into the environment
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Results
EEs of alpha-HCH, trans-,cis-chlordane in air samples and seawater samples range from 0.1 to 15.9% and 0.5 to 38.4%, respectively.EEs of metabolites heptachlor exo-epoxide in air samples and seawater samples range from 7.6 to 43.8% and 6.5 to 38.8%, respectively.The range of EEs of metabolites o,p’-DDD in seawater samples were from 20.7 to 64.4%.
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54.0 54.4 54.8 55.2 55.6 56.0 56.4 56.8 57.2Retention Time (min)
(1003)
(14412)
0
25
50
75
100
Inte
nsi
ty
chlordane / Average
54.4 54.8 55.2 55.6 56.0 56.4 56.8 57.2 57.6Retention Time (min)
(2054268)
0
25
50
75
100
Inte
nsi
tychlordane / Average
56.8 57.2 57.6 58.0 58.4 58.8 59.2 59.6 60.0Retention Time (min)
(521)
(4236)
0
25
50
75
100
Inte
nsi
ty
chlordane / Average
57.2 57.6 58.0 58.4 58.8 59.2 59.6 60.0 60.4Retention Time (min)
(535)
(9319)
0
25
50
75
100
Inte
nsi
ty
chlordane / Average
trans-
cis-
MC5
EE= 5 %
EE= 5 %
EE= 7 %
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S-1S-2
S-3
S-4S-5
S-6S-7
S-8S-9
S-10
S-11
EE values (South China Sea)
unit : %
from toalpha-HCH 6.3 1.3 15 3.6 o,p'-DDD 50 34 64 11 heptachlor exo-epoxide 39 39 39 -heptachlor endo-epoxide - - - -trans - chlordane 4.4 0.9 9.0 2.0 cis - chlordane 5.1 0.5 14 3.6 oxychlordane - - - -
average range standarddeviation
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P-1
P-2
P-3
P-4
P-5 P-7
P-6
P-8
P-9
P-10
P-11
EE values (Pacific Ocean)
unit : %
from toalpha-HCH 7.6 2.2 13 3.7 o,p'-DDD 56 21 70 18 heptachlor exo-epoxide 21 6.5 30 8 heptachlor endo-epoxide - - - -trans - chlordane 7.6 0.6 38 10.5 cis - chlordane 6.5 1.0 19 5.2 oxychlordane - - - -
average range standarddeviation
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N-1
N-2
N-3
N-4
EE values (North Atlantic Ocean)
unit : %
from toalpha-HCH 2.1 0.1 4.5 2.0 o,p'-DDD - - - -heptachlor exo-epoxide 31 7.6 44 16 heptachlor endo-epoxide - - - -trans - chlordane 6.2 0.4 16 6.7 cis - chlordane 3.4 2.0 7.1 2.5 oxychlordane 3.5 1.4 5.7 3.0
rangeaverage standarddeviation