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

IntroductionIntroduction

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.

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.

Materialsand

Methods

Materialsand

Methods

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 ～

P-1

P-2

P-3

P-4

P-5

P-6

P-7

Sampling Sites (Seawater)～ Pacific Ocean (West) ～

August～ September 2005

P-8

P-9

P-10P-11

August～ September 2005

UNITED STATES

San Francisco

Sampling Sites (Seawater)～ Pacific Ocean (East) ～

N-1

N-2

N-3

N-4

May～ June 2005

Sampling Sites (Air)～ North Atlantic Ocean ～

Marine pollution observation systemMarine pollution observation system

Solid PhaseExtraction unit

Control Unit

ExtractionColumn

Filter unit

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

Target Analytes

trans - Chlordane

Heptachlor α - HCH

Heptachlor epoxideOxychlordane

o,p’ - DDT

o,p’ - DDE

cis - Chlordane

Technical compounds

o,p’ - DDD

Metabolites

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 ）

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

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 %

ResultsResults

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

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)

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

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

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

Seawater (South China Sea)

0

20

40

60

80

o,p'-DDD heptachlorexo-epoxide

alpha-HCH trans-chlordane

cis-chlordane

EE(%)

Metabolite Technical compound

Seawater (Pacific Ocean)

0

20

40

60

80

o,p'-DDD heptachlorexo-epoxide

alpha-HCH trans-chlordane

cis-chlordane

EE(%)

Metabolite Technical compound

Air (North Atlantic Ocean)

0

25

50

heptachlorexo-epoxide

oxychlordane alpha-HCH trans-chlordane

cis-chlordane

EE(%)

Metabolite Technical compound

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.

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.

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

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

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

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

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

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

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

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%.

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 %

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

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

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