Andrew Loh1,2, Un Hyuk Yim1,2, Sung Yong Ha1 , Joon Geon An1 and Won Joon Shim1,2

1Korea Institute of Ocean Science and Technology (KIOST)2University of Science and Technology, Korea (UST)

OSA as an unknown route of exposure to filter feeding bivalve in the turbid

environment

� Introduction

�Materials and method

�Results and Discussion

�Conclusion

�Q & A

Content

� The Hebei Spirit Oil Spill: The Largest Oil Spill in Korea

� Collision of MV Hebei Spirit with a barge on7th December 2007 released approximately12,547 kL (10,900 M/T) of crude oil.

� The MV Hebei Spirit was carrying three kindsof crude oil, namely UAE Upper Zakum (UZC),Kuwait export crude (KEC) and Iranian heavycrude (IHC).

� Due to the strong westerly wind, spilled oilpolluted most of west coast of Korea, 375 kmcoastlines.

� More than one million of volunteers joined forinitial oil cleanup.

� Oil spill compensation process is still underway.

Introduction

Seawater Bivalves Sediments

Multi-media monitoring for oil contamination

16 PAHs

2007 Dec

2008 Jan

2008 Feb

2008 Mar

2008 Apr

2008 May

2008 Jun

2008 Jul

2008 Aug

2008 Sep

2008 Oct

2008 Nov

2008 Dec

2009 Feb

2009 Apr

2009 Jun

2009 Sep

2009 Dec

2010 Mar

2010 Jun

2010 Sep

2010 Dec

2011 Mar

2011 Jun

2011 Sep

2011 Dec

2012 Apr

2013 Apr

16 P

AH

s in

Oyst

er (

ng

/g d

w.)

0

200

400

600

800

1000

1200

1400

Alkylated PAHs

2007 Dec

2008 Jan

2008 Feb

2008 Mar

2008 Apr

2008 May

2008 Jun

2008 Jul

2008 Aug

2008 Sep

2008 Oct

2008 Nov

2008 Dec

2009 Feb

2009 Apr

2009 Jun

2009 Sep

2009 Dec

2010 Mar

2010 Jun

2010 Sep

2010 Dec

2011 Mar

2011 Jun

2011 Sep

2011 Dec

2012 Apr

2013 Apr

Alk

yl

PA

Hs

in O

yst

er

(ng/g

dw

.)

100

101

102

103

104

105

106

GC/FID Chromatogram

OysterDec 2007

Oyster Jan 2008

Oyster Apr 2008

Long-Term Monitoring Oil Fingerprinting

Stranded Oil

Dec 2007

min5 10 15 20 25 30

pA

0

200

400

600

800

1000

1200

- Initial: oil droplet- Persistent source

- 16 PAHs decreased rapidly- Alkyl PAHs persisted

- PAH double ratio- Manila clam: same with oyster

Oyster

Clam

� Route of exposure using exposure media� Traditional route of exposures; Water Accommodated Fractions (WAF) and Mechanically

Dissolved Oil (MDO).� Traditional methods could not match the Petroleum Derived Hydrocarbon distributions in

oysters from spill site.� Tests of unknown mode of exposure; Oil SPM Aggregates (OSA)

� Research objectives� To compare the fate of oil in the traditionally used exposure

methods.� To identify the continuous source of oil contamination to bivalves

in the HSOS.� To identify the fate of spilled oil in high turbidity environments

such as intertidal areas.

� OSA formation as the output of oil and particle interactions

� When oil and suspended particles interact, OSA is formed.� OSA formation has been observed in several large spills;

Exxon Valdez (1989), Sea Empress (1996) and Deep WaterHorizon (2010).

� OSA is formed in 2 main steps;(1) Breaking of surface oil by wave actions(2) Interaction of oil and particles

10µm

A B

100µm

100 µm

C

100 µm

D

� Formation of WAF� Method following CROSERF with slight modification(1) 1 L filtered seawater + magnetic stirrer(2) Addition of 25 g of Iranian Heavy Crude Oil(3) Magnetic stirred at 120 rpm for 24 hours(4) 900 ml of solution excluding surface oil was collected

� Formation of MDO(1) 1 L filtered seawater placed in separatory funnel(2) Addition of 25 g of Iranian Heavy Crude Oil(3) Vertical shaking at 50 rpm for 15 minutes(4) Settled for 1 hour(5) 900 ml of solution excluding surface oil was collected

� Formation of OSA� Method following Khelifa et a., 2002 with slight modification.(1) 1 L filtered seawater + 200 mg of particle(2) Solution + 600 mg of Iranian Heavy Crude oil

(Approxi. 1/40 of MDO and WAF)(3) Reciprocal shaking for 24 hours(4) Solution settled for 24 hours(5) 900 ml of solution excluding surface oil was collected

Materials and Method

� Exposure of media to oysters� Prior to exposure, oysters were acclimated in filtered seawater for 3 days.� Oysters are placed into the beaker filled with 700 ml of exposure media and a small

magnetic stirrer.� 10-AU fluorometer equipped with oil kit was used to quantify total petroleum

hydrocarbons before and after exposure.� After 24 hours of exposure, exposure media was replaced with newly prepared

exposure media.� After 48 hours of exposure, oysters were collected and prepared for chemical analysis

using GC/FID and GC/MS.

MDO OSA WAF

10-AU Fluorometer GC-MS GC-FID

Acclimation

MDO

WAF

Results and Discussion

OSA

Visual observation of oyster uptake

Exposure mediaBefore exposure

(ppm)After 24 hrs exposure

(ppm)Total loss (ppm)

MDO 238.0 109.0 129.0 (highest)

OSA 48.0 44.4 4.4 (middle)

WAF 0.92 0.67 0.25 (lowest)

Befo

re e

xposu

re

After

exp

osu

re

MDO

OSA

WAF

TPH Changes of Exposure Media

MDO

OSA

WAF

GC/FID chromatograms of petroleum hydrocarbons in oysters

Control

MDO

WAF

OSA

� TPH, UCM, n-alkane and Alkyl PAHs have concentration trend;MDO>OSA>WAF.

� But n-alkane for WAF exposed oyster was similar with control oyster.

� 16 PAHs have similar concentrations for all oysters, however, theconcentration of Alkylated PAHs was highest from MDO>OSA>WAF.

� This shows that, monitoring 16 PAHs alone is not suitable.

� Monitoring Alkylated PAHs could provide more accurate evaluation.

Control MDO (highest)

OSA

(middle)

WAF (lowest)

TPH 18.5 113.0 ± 52.0 66.2 ± 15.9 35.8 ± 6.9

UCM 8.7 66.9 ± 39.9 28.1 ± 10.6 12.7 ± 0.6

n-Alkane 0.5 7.1 ± 6.3 1.1 ± 0.6 0.3 ± 0.0

16 PAHs 0.1 3.0 ± 0.5 2.8 ± 0.6 2.4 ± 0.9

Alkyl

PAHs

0.360.2 ± 19.9

35.2 ± 6.113.8 ± 1.8

Accumulation of Petroleum Derived Hydrocarbons in oysters

Comparison of PAH profiles

CN

0

CN

1

CN

2

CN

3

CN

4

Acn

l

Acn

t

C0

F

C1

F

C2

F

C3

F

C0

P

An

th

C1p

C2

P

C3

P

C4

P

C0

D

C1

D

C2

D

C3

D

Flr

t

Py

r

Ba

A

C0

C

C1

C

C2

C

C3

C

Bb

F

Bk

F

Ba

P

Icd

P

Da

hA

Bg

hiPP

AH

s C

om

posi

tio

n i

n o

yst

ers

at

Sp

ill

Sit

e (

%)

0

5

10

15

20

25

30

Oyster Dec. 2007

CN

0

CN

1

CN

2

CN

3

CN

4

Acn

l

Acn

t

C0

F

C1

F

C2

F

C3

F

C0

P

An

th

C1

p

C2

P

C3

P

C4

P

C0D

C1D

C2D

C3D

Flr

t

Py

r

BaA

C0C

C1C

C2C

C3C

Bb

F

Bk

F

BaP

Icd

P

Da

hA

Bgh

iPPA

Hs

Co

mp

osi

tion

in

oy

ster

s at

Sp

ill

Sit

e (%

)

0

10

20

30

40

50

Oyster Dec. 2008

CN

0

CN

1

CN

2

CN

3

CN

4

Acn

l

Acn

t

C0

F

C1

F

C2

F

C3

F

C0

P

An

th

C1p

C2

P

C3

P

C4

P

C0

D

C1

D

C2

D

C3

D

Flr

t

Pyr

Ba

A

C0

C

C1

C

C2

C

C3

C

Bb

F

Bk

F

Ba

P

Icd

P

Da

hA

Bg

hiPPA

Hs

Co

mp

osi

tion

in

sea

wate

r o

f sp

ill

site

(%

)

0

5

10

15

20

25

30

Seawater Dec. 2007

CN

0

CN

1

CN

2

CN

3

CN

4

Acn

l

Acn

t

C0

F

C1

F

C2

F

C3

F

C0

P

An

th

C1p

C2

P

C3

P

C4

P

C0

D

C1

D

C2

D

C3

D

Flr

t

Pyr

Ba

A

C0

C

C1

C

C2

C

C3

C

Bb

F

Bk

F

Ba

P

Icd

P

Da

hA

Bg

hiPPA

Hs

Co

mp

osi

tion

in

Ira

nia

n H

eav

y C

rud

e (%

)

0

5

10

15

20

25

30

Iranian Heavy Crude

Field Samples

Exposure Media Oysters

OSA

C0

NC

1N

C2

NC

3N

C4

NB

iPA

cnl

Acnt

C0

FC

1F

C2

FC

3F

C0

PA

nth

C1

PC

2P

C3

PC

4P

C0

DC

1D

C2

DC

3D

Flr

tP

yr

BaA

C0C

C1C

C2C

C3C

Bb

FB

kF

BeP

BaP

Icd

PD

ahA

Bg

hiP

PerP

AH

Co

mp

osi

tio

n i

n O

yst

er

(%)

0

5

10

15

20

25

30

WAF

C0N

C1N

C2N

C3N

C4N

BiP

Acn

lA

cnt

C0

FC

1F

C2

FC

3F

C0

PA

nth

C1

PC

2P

C3

PC

4P

C0D

C1D

C2D

C3D

Flr

tP

yr

BaA

C0C

C1C

C2C

C3C

Bb

FB

kF

BeP

BaP

Icd

PD

ahA

Bg

hiP

Per

PA

H C

om

po

siti

on

in

Oy

ster

(%)

0

10

20

30

40

50

MDO

C0N

C1N

C2N

C3N

C4N

BiP

Acn

lA

cnt

C0

FC

1F

C2

FC

3F

C0

PA

nth

C1

PC

2P

C3

PC

4P

C0D

C1D

C2D

C3D

Flr

tP

yr

BaA

C0C

C1C

C2C

C3C

Bb

FB

kF

BeP

BaP

Icd

PD

ahA

Bg

hiP

PerP

AH

Co

mp

osi

tio

n i

n O

yst

er (

%)

0

5

10

15

20

25

WAF

C0

NC

1N

C2

NC

3N

C4

NB

iPA

cnl

Acn

tC

0F

C1F

C2F

C3F

C0P

Anth

C1P

C2P

C3P

C4P

C0

DC

1D

C2

DC

3D

Flr

tP

yr

BaA

C0

CC

1C

C2

CC

3C

BbF

BkF

BeP

BaP

IcdP

Dah

AB

gh

iPP

erPA

H C

om

po

siti

on

in

Sea

wa

ter (

%)

0

10

20

30

40

50

MDO

C0

NC

1N

C2

NC

3N

C4

NB

iPA

cnl

Acn

tC

0F

C1F

C2F

C3F

C0P

Anth

C1P

C2P

C3P

C4P

C0

DC

1D

C2

DC

3D

Flr

tP

yr

BaA

C0

CC

1C

C2

CC

3C

BbF

BkF

BeP

BaP

IcdP

Dah

AB

gh

iPP

er

PA

H C

om

po

siti

on

in

Seaw

ate

r (%

)

0

10

20

30

40

OSA

C0N

C1N

C2N

C3N

C4N

BiP

Acn

lA

cnt

C0

FC

1F

C2

FC

3F

C0

PA

nth

C1

PC

2P

C3

PC

4P

C0D

C1D

C2D

C3D

Flr

tP

yr

BaA

C0C

C1C

C2C

C3C

Bb

FB

kF

BeP

BaP

Icd

PD

ahA

Bg

hiP

PerPA

H C

om

posi

tio

n i

n S

eaw

ate

r (%

)

0

10

20

30

40

50

60

Y D

ata

Y D

ata

Y D

ata

Route of Exposure Tests

노노노

ControlWAF OSA MDO

16

PA

Hs

Co

nc.

in

Oy

ster

(n

g/g

dw

.)

100

101

102

103

104

105

106

노노노

ControlWAF OSA MDO

Alk

yl

PA

Hs

Co

nc.

in

Oy

ster

(n

g/g

dw

.)

100

101

102

103

104

105

106

PAH double ratio plots

C2D/C2P ratio

0.0 0.5 1.0 1.5 2.0 2.5 3.0

C3

D/C

3P

ra

tio

0.0

0.5

1.0

1.5

2.0

2.5

3.0 IHC(SGS)

ControlWAF1WAF2

WAF3

OSA1OSA2OSA3

MDO1

MDO2

MDO3

� 16 PAHs concentrations could not distinguish their route of exposure but alkyl PAHscould.

� Alkyl PAHs could be used in double ratio plots to identify their relativity with source oil.� Double ratios using alkylated Phenanthrene and alkylated Dibenzothiophene has been

widely used for oil fingerprinting.� Double ratios of MDO and OSA matched more with the source oil but WAF did not

match.� Double ratio plots are very useful fingerprinting tool.

WAF OSA MDO

� To explain persistency of PAHs in oysters after the Hebei Spirit oil spill,available exposure method including WAF, MDO, and OSA were tested.

� Among three exposure media, OSA showed higher accumulationefficiency than others.

� Alkylated PAHs in oyster well exhibited accumulation of petroleumderived hydrocarbons.

� PAHs double ratio in oyster was proved to be useful for oilfingerprinting.

� OSA could be used as a new route of exposure to study thebioaccumulation of oil.

Conclusion

End of Presentation