Kansas State University - clu-in.org · 4 Kulakow, Int. App. Phytotechnologies Conf., 5 March 2003 • Experimental Design – Randomized complete block design with four replications

1

Kulakow, Int. App. Phytotechnologies Conf., 5 March 2003

RTDF TPH Subgroup Phytoremediation Field Trials:

Results and Preliminary Conclusions

Peter Kulakow and Xiujuan FengKansas State University


Acknowledgements

• RTDF Cooperators under the leadership of– Steve Rock– Lucinda Jackson– Phil Sayre

• USEPA Technology Innovation Office• Environment Canada• Great/Plains Rocky Mountain HSRC• Midwest HSRC• Henry Camp - ICF Consulting

2


RTDF Partners

• US Environmental Protection Agency• Environment Canada• Major Oil Corporations

– Petroleum Environmental Research Forum• Public Utility Corporations• US Department of Defense• Universities• Environmental Consultants


ACCORDING TO: www.epa.gov/greenacres/8

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Root Systems of Prairie Plants - From the U.S. EPA Handbook on Natural Landscapes

KentuckyBluegrass

LeadPlant

MissouriGoldenrod

IndianGrass

CompassPlant

PorcupineGrass

HeathAster

PrairieCordgrass

Big BlueStem

PalePurple

Coneflower

PrairieDropseed

Side OatsGrama

FalseBoneset

SwitchGrass

WhiteWild Indigo

LittleBlue Stem

RosinWeed

PurplePrairieClover

JuneGrass

CylindricBlazing

Star

BuffaloGrass 15 ft

TypicalLawnGrass

EfficientRoot

Producer

Deep-Rooted Species 4

3


Goal

• Assess the efficacy of vegetation to enhance degradation of aged petroleum hydrocarbons in soil

• Is there evidence that vegetation enhances progress toward practical environmental management objectives for petroleum sites within 3 years?


Rationale for Approach

• Hydrocarbon degradation – Changes are slow and subtle

• Monitoring is needed for a long period• Contaminant distribution in soil is

variable• Standard protocol facilitates

comparisons among locations – allowing for site specific adjustments

4


• Experimental Design– Randomized complete block design with

four replications– Treatments with site specific adjustments

–Standardized grass/legume mixture–Local-optimized treatment–Unvegetated/unfertilized Control

– Minimum plot size: 20 x 20 ft.

Standardized Protocol


Site I

110 ft

185 ft20 ft100 ft

X

35 ft

70 ft

140 ft

125 ft

225 ft

HEAVIERFORESTED

AREA100 ft

PONDED AREA

ROADWAYBERM

AccessRamp

Hydrant

SurfacePipe

SurfacePipe

ROADWAYBERM

ROADWAYBERM

A

C

A

B

CD

A

B

C

D

A

BC

DSTAGING AREA

FOR FIELDACTIVITIES

BD

REVISEDLAYOUT

GravellyArea

RTDF: 40 lbs fescue / 10 lbs p.rye / 5 lbs cloverPrairie: 10 lbs prairie / 25 lbs fescue

A = Do NothingB = Fertilizer OnlyC = RTDF MixD = Prairie Mix

5


• Soil– Composite 8 soil cores/plot – Two depths: 0 – 15 cm and 15 – 45 cm– Sample Annually for 3 growing seasons

• Vegetation– Above ground biomass production – Plant species composition – Root growth– Hydrocarbon uptake at final sampling

Sampling Plan


Site A – May 2001Hydrocarbon Uptake Sampling

6


Sample Preparation and Analysis Flow

10 grams of sample is Soxhlet

extracted

OIL & GREASE (GRAV)OIL & GREASE (GRAV)

Weight measured

gravimetrically

Extract column cleanup

Weight measured

gravimetricallySplit for

instrument analyses

TPH (GRAV)TPH (GRAV)

TPH (GC/FID)TPH (GC/FID)

PAH (GC/MS)PAH (GC/MS)

HOPANE (GC/MS)HOPANE (GC/MS)


Example GC/FID Chromatogram Output

0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00

1000

2000

3000

4000

5000

6000

7000

8000

9000

10000

11000

12000

13000

14000

Time

Response_

03070025.D\FID2B

Resolved Components (TPH RES)

Unresolved Components (TPH)

7


0

500

1000

1500

2000

2500

3000

Nap

htha

lene

C1-

Nap

htha

lene

s

C2-

Nap

htha

lene

s

C3-

Nap

htha

lene

s

C4-

Nap

htha

lene

s

Acen

apht

hyle

ne

Acen

apht

hene

Biph

enyl

Fluo

rene

C1-

Fluo

rene

s

C2-

Fluo

rene

s

C3-

Fluo

rene

s

Anth

race

ne

Phen

anth

rene

C1-

Phen

anth

rene

s/an

thra

cene

s

C2-

Phen

anth

rene

s/an

thra

cene

s

C3-

Phen

anth

rene

s/an

thra

cene

s

C4-

Phen

anth

rene

s/an

thra

cene

s

Dib

enzo

thio

phen

e

C1-

Dib

enzo

thio

phen

es

C2-

Dib

enzo

thio

phen

es

C3-

Dib

enzo

thio

phen

es

Fluo

rant

hene

Pyre

ne

C1-

Fluo

rant

hene

s/py

rene

s

C2-

Fluo

rant

hene

s/py

rene

s

C3-

Fluo

rant

hene

s/py

rene

s

Benz

o[a]

anth

race

ne

Chr

ysen

e

C1-

Chr

ysen

es

C2-

Chr

ysen

es

C3-

Chr

ysen

es

C4-

Chr

ysen

es

Benz

o[b]

fluor

anth

ene

Benz

o[k]

fluor

anth

ene

Benz

o[e]

pyre

ne

Benz

o[a]

pyre

ne

Pery

lene

Inde

no[1

,2,3

,-c,d

]pyr

ene

Dib

enzo

[a,h

]ant

hrac

ene

Benz

o[g,

h,i]p

eryl

ene

C om poun ds

Con

cent

ratio

n ug

/Kg

" 20A 4455: G 11T 1A S O IL "Analyte P ro file H istogram

Example Expanded PAH OutputPAH Homologous

SeriesSulfur-containing

Heterocyclics 5- and 6-Ring PAH


TPHCWG Sample Preparation and Analysis Flow

10 grams of sample is

vortex extracted in

pentane

Extract is fractionated and cleaned

Each fraction submitted for

instrument analyses

Aliphatic TPH Ranges (9)Aliphatic TPH Ranges (9)

Aromatic TPH Ranges (9)Aromatic TPH Ranges (9)

8


Example TPHCWG Chromatogram Output

0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00

0

5000

10000

15000

20000

25000

30000

35000

40000

45000

50000

55000

60000

65000

70000

75000

80000

85000

90000

95000

Time

Response_

08300121.D\FID2B

0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00

0

5000

10000

15000

20000

25000

30000

35000

40000

45000

50000

55000

Time

Response_

08300122.D\FID1A

Quantitation divided in ranges related to

boiling point


TPH-CWGM

Total Resolved 210Total PHC 2000

Aliphatic Total Resolved 180Aliphatic TPH 1500Aliphatic TPH>nC6-nC8 NDAliphatic TPH>nC8-nC10 1.3 JAliphatic TPH>nC10-nC12 10 JAliphatic TPH>nC12-nC16 120Aliphatic TPH>nC16-nC21 210Aliphatic TPH>nC21-nC25 240Aliphatic TPH>nC25-nC30 500Aliphatic TPH>nC30-nC35 280Aliphatic TPH>nC35+ 140

Aromatic Total Resolved 33Aromatic TPH 480Aromatic TPH>nC6-nC7 (B) NDAromatic TPH>nC7-nC8 (T) NDAromatic TPH>nC8-nC10 NDAromatic TPH>nC10-nC12 0.15 JAromatic TPH>nC12-nC16 26Aromatic TPH>nC16-nC21 90Aromatic TPH>nC21-nC25 74Aromatic TPH>nC25-nC30 120Aromatic TPH>nC30-nC35 98Aromatic TPH>nC35+ 70

TPH-CWGM

Total Resolved 210Total PHC 2000

Aliphatic Total Resolved 180Aliphatic TPH 1500Aliphatic TPH>nC6-nC8 NDAliphatic TPH>nC8-nC10 1.3 JAliphatic TPH>nC10-nC12 10 JAliphatic TPH>nC12-nC16 120Aliphatic TPH>nC16-nC21 210Aliphatic TPH>nC21-nC25 240Aliphatic TPH>nC25-nC30 500Aliphatic TPH>nC30-nC35 280Aliphatic TPH>nC35+ 140

Aromatic Total Resolved 33Aromatic TPH 480Aromatic TPH>nC6-nC7 (B) NDAromatic TPH>nC7-nC8 (T) NDAromatic TPH>nC8-nC10 NDAromatic TPH>nC10-nC12 0.15 JAromatic TPH>nC12-nC16 26Aromatic TPH>nC16-nC21 90Aromatic TPH>nC21-nC25 74Aromatic TPH>nC25-nC30 120Aromatic TPH>nC30-nC35 98Aromatic TPH>nC35+ 70

0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00

0

5000

10000

15000

20000

25000

30000

35000

40000

45000

50000

55000

60000

65000

70000

75000

80000

85000

90000

95000

Time

Response_

08300121.D\FID2B

0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00

0

5000

10000

15000

20000

25000

30000

35000

40000

45000

50000

55000

Time

Response_

08300122.D\FID1A

TPHCWGData

9


Thirteen RTDF Field Site Locations


April 1999

May 2001June 2000

Site A – Refinery Site

Three Complete Growing Seasons

10


Site B - Closed RefineryRTDF Mix

Poplar/Willow Hackberry

Unvegetated


1/1/98 4/1/98

7/1/98

10/1/98 1/1/99

4/1/99 7/1/99

10/1/99 1/1/00

4/1/00 7/1/00

10/1/00 1/1/01

4/1/01 7/1/01

10/1/01 1/1/02

4/1/02 7/1/02

10/1/02 1/1/03

-10

-5

0

5

10

15

20

25

30

-15

-10

-5

0

5

10

15

20

25

30

35

40

Tem

p era

ture

(oC

)

Pre

cipi

tatio

n (c

m)

Time (month)

Site B, OH (1998Site B, OH (1998--2002)2002)

Planting

11


Three Alaskan Sites – Cold Regions

Site C Site E

Site D


1/1/98 4/1/98

7/1/98

10/1/98 1/1/99

4/1/99 7/1/99

10/1/99 1/1/00

4/1/00 7/1/00

10/1/00 1/1/01

4/1/01 7/1/01

10/1/01 1/1/02

4/1/02 7/1/02

10/1/02 1/1/03

-20

-15

-10

-5

0

5

10

-35

-30

-25

-20

-15

-10

-5

0

5

10

15

Tem

p era

ture

(oC

)

Pre

cipi

tatio

n (c

m)

Time (month)

Site C (1998Site C (1998--2002)2002)

TMax

TMin

Pcp

Planting

12


1/1/98 4/1/98

7/1/98

10/1/98 1/1/99

4/1/99 7/1/99

10/1/99 1/1/00

4/1/00 7/1/00

10/1/00 1/1/01

4/1/01 7/1/01

10/1/01 1/1/02

4/1/02 7/1/02

10/1/02 1/1/03

-5

0

5

10

15

20

25

30

35

40

45

-5

0

5

10

15

20

25

Time (month)

Pre

cipi

tatio

n (c

m)

Tem

p era

ture

(oC

)TMax

TMin

Pcp

Site E (1998Site E (1998--2002)2002)

Planting


Site F – Manufactured Gas PlantWillow/Poplar Mix

RTDF Mix

13


Willow Willow

Willow

Willow

Grass

Grass

Grass

Grass

Volunteer

Volunteer

Volunteer

Volunteer

NoVegetation

NoVegetation

NoVegetation

No Vegetation

North

Water tank

Gravel road

Gate

Rep. 1 Rep. 2

Rep. 3Rep. 4

Site F – New York


Site G – Kansas – Motor Pool Waste Lagoon Sediments

14


1/1/98 4/1/98

7/1/98

10/1/98 1/1/99

4/1/99 7/1/99

10/1/99 1/1/00

4/1/00 7/1/00

10/1/00 1/1/01

4/1/01 7/1/01

10/1/01 1/1/02

4/1/02 7/1/02

10/1/02 1/1/03

-5

0

5

10

15

20

25

-15

-10

-5

0

5

10

15

20

25

30

35

40Te

mp e

ratu

re (o

C)

Pre

cipi

tatio

n (c

m)

Time (month)

Site G (1998Site G (1998--2002)2002)

TMax

TMin

Pcp

Planting


Site I – Closed RefineryIllinois

October 2001 May 2002

15


Site H – Closed Distribution FacilityRhode IslandOctober 2002


Site J – Production SiteArkansas

16


Site K – Manufactured Gas Plant, Indiana


Site L – SK, Canada – Buried Flare Pit

Planted June 2002

17


Results


Status of Field Sites – 36 Data Sets

Oct-05May-03May-03Site M, AL Canada

Oct-04Oct-02Jun-02Jun-02Site L, SK Canada

May-99Site K, IN

May-02Oct-01Oct-99Site J, AR

Nov-03Oct-02Nov-00Site I, MO

Nov-03Oct-02May-01May-01Site H, RI

Oct-02Oct-99Site G, KS

Oct-02Jun-99Site F, NY

Aug-98Site E, AK

Aug-98Aug-98Site D, AK

Jun-99Site C, AK

Nov-02Apr-99Site B, OH

Dec-98Site A, CA

Year 4Year 3Year 2Year 1Time 0Date

Sampling EventsPlanting

18


Dealing with the Data

• Nondetect data– Used a nonzero substitution

• Numerous derived variables such as Total PAHs– >100 variables analyzed

• Analyzed– Original data– Hopane normalized data


Statistical Analysis

• Randomized Complete Block Design• Soil depth usually analyzed separately• Time is a repeated measure

– Derived variables approach– Each time interval analyzed separately

• Considered assumptions of analysis of variance– Homogeneity of variances most important

• Treatment differences tested by analysis of variance and lsd tests.

19


x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

Site A

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

StandardReferenceSampleSite A

Tota

l PA

Hs

mg/

Kg

95% Confidence Ellipses for RTDF Data


High Variability in TPH and PAH Concentrations• Between locations

– Original composition of contaminants– Previous degradation– Climate

• Within locations– Spatial variation

• some accounting for in blocking of experiments– Time, Depth, and Treatment effects– Management Decisions

• Within samples– Sample heterogeneity– Analytical Variation

• Instrumentation• Laboratory Procedures

20


x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000


x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

Site A

Site B

TPH (mg/Kg)

TPH (mg/Kg)

x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

Site GTo

tal P

AH

sm

g/K

g

x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

Site F

TPH (mg/Kg)

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

Tota

l PA

Hs

mg/

Kg

Tota

l PA

Hs

mg/

Kg


95% Confidence Ellipses for RTDF Datax2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

Site D

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

Site C

Site E

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

21


x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

Site H

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg


x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

Site I

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

Site J

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

x2

-1500

-900

-300

300

900

1500

2100

2700

3300

3900

4500

-20000 0 20000 40000 60000 80000 100000 120000

TPH (mg/Kg)

Tota

l PA

Hs

mg/

Kg

StandardReferenceSample


Site F Treatment Means for Total PAHs for 0 to 15 cm Depth

0

50

100

150

200

250

300

350

400

450

500

0 6 18 30Months

Con

cent

ratio

n (m

g/kg

)

Volunteer RTDF Mixture Unvegetated Willow/Poplar

22


Site F PAH Treatment Means18 Months, Depth 0 to 15 cm

0

5

10

15

20

25C

0N

C1N

C2N

C3N

C4N

AC

EY

AC

E

BIP

C0F

C1F

C2F

C3F

C0A

C0P

C1P

_A

C2P

_A

C3P

_A

C4P

_A

C0D

C1D

C2D

C3D

FLA

NT

PY

R

C1F

_P

C2F

_P

C3F

_P BA

A

C0C

C1C

C2C

C3C

C4C BBF

BKF

BE

P

BAP

PER

IND

EN

O

DA

H

BGP

PAH

Con

cent

ratio

n (m

g/kg

)

Volunteer Standard Unvegetated Willow


Treatment Differences for 41 PAHs>25% of PAHs different by ANOVA (P<0.05)

Site MSite LSite K

Site JSite ISite H

0.630.390.510.44Site G0.61Site F

Site ESite DSite CSite B

0.290.24Site ABA BA BA BA Depth

Year 3Year 2Year 1Time 0Sampling Events

23


Treatment Differences for 41 Normalized PAHs>25% of PAHs different by ANOVA (P<0.05)

Site MSite LSite KSite JSite ISite H

0.800.560.710.46Site GSite FSite ESite DSite CSite B

0.420.540.39Site ABA BA BA BA Depth

Year 3Year 2Year 1Time 0Sampling Events


0

5

10

15

20

25

30

35

40

45

50

0 12 24Months

PAH

to H

opan

e R

atio

Native Standard Unvegetated

Site G – Hopane Normalized Total PAHs0 – 15 cm Depth

T1-T0 T2-T0

24

Kulakow, Int. App. Phytotechnologies Conf., 5 March 2003Site MSite LSite KSite JSite ISite H

0.510.460.440.44Site G0.34Site F

Site E0.53Site D

Site CSite B

0.56.49.42.370.390.29Site A

BA BA BA BA Depth

Time 3-Time 1Time 3-Time 0Time 2-Time 0Time 1-Time 0Interval

Treatment DifferencesChange in 41 Normalized PAHs

>25% of PAHs different by ANOVA (P<0.05)


0

10

20

30

40

50

60

70

80

0 12 24Months

PAH

to H

opan

e R

atio


Site G – Hopane Normalized Total PAHs15 – 45 cm Depth

25


Total Petroleum Hydrocarbons -- Plot G11, 0-15 cm Depth

0-months

24-months

12-months

TPH13,000 mg/kg

9,500 mg/kg

3,900 mg/kg


AliphaticAromatic

Time 0

Site G – 0 to 15 cm Depth

No Vegetation24 months

RTDF MixVegetation24 months

22%

78%

23%

77%

30%

70%

26


C6-8C8-10C10-12C12-16C16-21C21-35C35+

Aliphatic Fractions Time 0



RTDF MixVegetation24 months

77%70%

62%

70%

14%

11%9%

8%13%12%

74%

10% 7%

3%


C8-10C10--12C12-16C16-21C21-35C35+

Aromatic Fractions Time 0



RTDF Mix24 months

77%70%

5%

14%17% 15%

69% 60%

20%

24%46%

7%

17%

27


RTDF Trial Site A -- Hopane Normalized Total PAHs 15 to 45 cm

0

50

100

150

200

250

0 6 18 30Months

PAH

to H

opan

e R

atio



RTDF Trial Site A -- Hopane Normalized Total PAHs 0 to 15 cm

0

20

40

60

80

100

120

140

0 6 18 30Months

PAH

to H

opan

e R

atio


28


RTDF Trial Site J -- Total Petroleum Hydrocarbons0 to 15 cm

02000400060008000

10000120001400016000

0 9 19Months

Con

cent

ratio

n (m

g/kg

)

Bermudagrass Standard Unvegetated


Status of Project

• 75% data available• 4 new datasets within next month• 7 of 13 locations are finished

– 5 have data for 3 growing seasons • Site F will continue for 3 more years

– less than 3 growing seasons for 2 sites• Additional results expected for 7 locations• 2 new locations in Canada• More data analysis

29


Primary Results to Date

– evidence of negative vegetation effectin surface soil only

A

– evidence of emerging vegetation effect- Needs confirmation over longer period

A,D,F

– strong evidence of positive vegetationeffect only with an alternativesampling method

J

– strong evidence of positive vegetationeffect- Highly degradable source, lower variation

GEvidenceSites


Preliminary Conclusions

• Is there evidence that vegetation enhances progress toward practical environmental management objectives for petroleum sites within 3 years?

• Yes, at some locations. • Use site specific

30



• Best locations -- Highly degradable source material with low to moderate hydrocarbon concentrations near risk-based levels.

– Site characterization important– Net environmental benefits and cost may

support use of phytotechnologies.


Preliminary Conclusions• Highly weathered sites

– Evidence of vegetation effect difficult to demonstrate.

– High variability from multiple sources obscures potential treatment effects.

– Initial composition and aging limits potential for further degradation.

– Effects are long term and three years not always sufficient.

– Vegetation has both positive and negative effects on bioavailability.

31



• Highly weathered sites– Phytotechnologies may be part of a long

term ecological restoration strategy.– Phytorestoration that includes continued

slow remediation and stabilization processes.

– Need to understand bioavailability and risk.


The Future

• Improvements to the protocol• Additional results and analyses• Final report• Results on RTDF website

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TPH In Soil Subgroup

www.rtdf.org

Documents

Kansas State University - clu-in.org · 4 Kulakow, Int. App. Phytotechnologies Conf., 5 March 2003 • Experimental Design – Randomized complete block design with four replications