(201 3/2/1 Nagoya) - 大阪府立大学matsu/data/isplasma5/...3 Phorbol Esters Phorbol Esters Introduction Introduction Introduction Introduction J. Curcas J. Curcas seed seed kernel

Degradation of phorbol esters Degradation of phorbol esters Degradation of phorbol esters

Degradation of phorbol esters

with an atmospheric pressure with an atmospheric pressure with an atmospheric pressure

with an atmospheric pressure

plasma jetplasma jetplasma jet

plasma jet

ISPlasma 2013(2013/2/1、Nagoya)

Osaka Prefecture UniversityH.Matsuura, S.Kongmany

M.Furuta, K.Imamura,Y.Maeda and S.Okuda

Jatropha curcas (J. curcas) currently is the best candidate as the raw material for producing biodiesel since it contains lots of oil of approximately 60%.

But the Jatropha oil produced contains the toxic components of phorbolesters that act a cancer promoter. Nowadays, the solution for solving this problems by various physical and chemical means is investigating all over the world.

Sunlight(UV?)Ultrasonic waveGamma-ray

Atmospheric pressure plasma is also found to provide reactive free radicals to convert the phorbol ester into original phorbol. We now start measurement radical concentration in the solution to study the mechanism of degradation and to compare the efficiency of degradation methods.

Background

3

Phorbol EstersPhorbol EstersPhorbol Esters

Phorbol Esters

IntroductionIntroductionIntroduction

Introduction

IntroductionIntroductionIntroduction

Introduction

J. Curcas J. Curcas J. Curcas

J. Curcas

seed seed seed

seed

kernelkernelkernel

kernel

Mechanical Mechanical Mechanical

Mechanical

extraction of extraction of extraction of

extraction of

oiloiloil

oil

Biodiesel Biodiesel Biodiesel

Biodiesel

productionproductionproduction

production

Livestock Livestock Livestock

Livestock

feedingfeedingfeeding

feeding

Crude J. curcas OilCrude J. curcas OilCrude J. curcas Oil

Crude J. curcas Oil

J. Curcas seed cakeJ. Curcas seed cakeJ. Curcas seed cake

J. Curcas seed cake

Containing PEs

Containing PEs

Containing PEs

Containing PEs

• Oil: 40-60%Oil: 40-60%Oil: 40-60%

Oil: 40-60%

• Proteins: 20-30%Proteins: 20-30%Proteins: 20-30%

Proteins: 20-30%

• phorbol ester (PEs)phorbol ester (PEs)phorbol ester (PEs)

phorbol ester (PEs)

: 0.8-3.3 mg/g.

• One of the exploitation of renewable sources of energy:One of the exploitation of renewable sources of energy:One of the exploitation of renewable sources of energy:

One of the exploitation of renewable sources of energy:

• The production of biodiesel via esterification or transesterification:The production of biodiesel via esterification or transesterification:The production of biodiesel via esterification or transesterification:

The production of biodiesel via esterification or transesterification:

• Edible vegetable oils and animal fats, etc.Edible vegetable oils and animal fats, etc.Edible vegetable oils and animal fats, etc.

Edible vegetable oils and animal fats, etc.

• Non-edible oils: Non-edible oils: Non-edible oils:

Non-edible oils:

Jatropha curcas oil (J. curcas oil), Jatropha curcas oil (J. curcas oil), Jatropha curcas oil (J. curcas oil),

Jatropha curcas oil (J. curcas oil),

etc.etc.etc.

etc.

• In 2015, 12.8 Mt of J. curcas oil will be produced (GEXST, 2008).In 2015, 12.8 Mt of J. curcas oil will be produced (GEXST, 2008).In 2015, 12.8 Mt of J. curcas oil will be produced (GEXST, 2008).

In 2015, 12.8 Mt of J. curcas oil will be produced (GEXST, 2008).

70-75% PEs70-75% PEs70-75% PEs

70-75% PEs

20-25% PEs20-25% PEs20-25% PEs

20-25% PEs

PlasmPlasmaa Degradation of phorbol estersDegradation of phorbol esters• Mixed 4 standard phorbol ester in methanol were exposed to plasma channel at

constant He flow and applied voltage. The plasma ionizing time was optimized.

Radiation Research CenterRadiation Research CenterRadiation Research Center

Radiation Research Center

Fig. 5 Fig. 5 Fig. 5

Fig. 5

Atmospheric pressure plasma jet for degradation of mixed 4 phorbol esters solution.



aaa

a

bbb

b

ccc

c

Fig. 11 Fig. 11 Fig. 11

Fig. 11

Degradation profile of mixed 4 phorbol esters before and after plasma ionization: (a) &(b) the comparison of HPLC/UV chromatogram and (c) concentration and degradation of phorbol esters.

• PDA: PDA: PDA:

PDA:

phorbol 12,13-diacetate, PDBu, PDBu, PDBu

, PDBu

: 12,13-dibutyrate,

• PDB: PDB: PDB:

PDB:

phorbol 12,13-dibenzoate, • PMA (TPA)PMA (TPA)PMA (TPA)

PMA (TPA)

: phorbol 12-myristate 13-acetate

Treatment of aqueous solution

HPLC/UV chromatograms of aqueous solution of: (a) HPLC/UV chromatograms of aqueous solution of: (a) HPLC/UV chromatograms of aqueous solution of: (a)

HPLC/UV chromatograms of aqueous solution of: (a)

TPA and (b) mixed jatropha phorbol esters (4 peaks each TPA and (b) mixed jatropha phorbol esters (4 peaks each TPA and (b) mixed jatropha phorbol esters (4 peaks each

TPA and (b) mixed jatropha phorbol esters (4 peaks each

peak corresponding to each type of jatropha factor) peak corresponding to each type of jatropha factor) peak corresponding to each type of jatropha factor)

peak corresponding to each type of jatropha factor)

before and after plasma irradiation for 15 min.before and after plasma irradiation for 15 min.before and after plasma irradiation for 15 min.

before and after plasma irradiation for 15 min.

Phorbol esters: Phorbol esters: Phorbol esters:

Phorbol esters:

Expected degradation productExpected degradation productExpected degradation product

Expected degradation product

National University of Laos, Faculty of Science, Department of ChemistryNational University of Laos, Faculty of Science, Department of ChemistryNational University of Laos, Faculty of Science, Department of Chemistry

National University of Laos, Faculty of Science, Department of Chemistry

• TheTheThe

The

phorbol ester phorbol ester phorbol ester

phorbol ester

may be converted into may be converted into may be converted into

may be converted into

original phorbol original phorbol original phorbol

original phorbol

since it has since it has since it has

since it has

ever taken place during the transesterification process of J. curcas oil ever taken place during the transesterification process of J. curcas oil ever taken place during the transesterification process of J. curcas oil

ever taken place during the transesterification process of J. curcas oil

with alkaline/methanol. [Y. Maeda (2012)]with alkaline/methanol. [Y. Maeda (2012)]with alkaline/methanol. [Y. Maeda (2012)]

with alkaline/methanol. [Y. Maeda (2012)]

8

OC

CH2OH

H

OH

H3C

O

H

OH

H3C

CH3CH3

H

CCH3

H3C(H2C)11H2CC

O

O

O

US / US / US /

US /

γ-ray-ray-ray

-ray

2 (•OH) 2 (•H)

TetraDecanoyl Phorbol-13-Acetae (TPA)TetraDecanoyl Phorbol-13-Acetae (TPA)TetraDecanoyl Phorbol-13-Acetae (TPA)

TetraDecanoyl Phorbol-13-Acetae (TPA)

Phorbol 12-myristate 13-acetate (PMA)Phorbol 12-myristate 13-acetate (PMA)Phorbol 12-myristate 13-acetate (PMA)

Phorbol 12-myristate 13-acetate (PMA)

HOHO

CH2OH

H

OH

H3C

O

H

OH

H3C

CH3CH3

phorbol

H

PhorbolPhorbolPhorbol

Phorbol

Conclusion

• By using UV Chromatogram, degradation of Phorbol ester with atmospheric plasma and is confirmed.

• Concentration of hydroxyl radicals (OH) is estimated with KI-colorimetric method, of which result does not contradict with Prof. Kanazawa's work with the similar plasma device and different measurement method.

Future works• Test using PE with much simpler structure

to study degradation mechanism

• Control plasma parameters and study the relation between OH concentration and degradation effect

• Direct confirm of detoxification with biological manners, not merely disappear of Phorbol Ester

Appendix

Why Why Why

Why

is J. curcas used for biodiesel production if it contains PEs?is J. curcas used for biodiesel production if it contains PEs?is J. curcas used for biodiesel production if it contains PEs?

is J. curcas used for biodiesel production if it contains PEs?

Oil yield: Jatropha vs other seed crops.Oil yield: Jatropha vs other seed crops.Oil yield: Jatropha vs other seed crops.

Oil yield: Jatropha vs other seed crops.

Sources: Sources: Sources:

Sources:

http://www.infinitysource.com/projects/jatropha.shtmlhttp://www.infinitysource.com/projects/jatropha.shtmlhttp://www.infinitysource.com/projects/jatropha.shtml

http://www.infinitysource.com/projects/jatropha.shtml

, accessed 26-05-2012 , accessed 26-05-2012 , accessed 26-05-2012

, accessed 26-05-2012

• Jatropha give Jatropha give Jatropha give

Jatropha give

very highest very highest very highest

very highest

oil production oil production oil production

oil production

and it is non-and it is non-and it is non-

and it is non-

edible oil.edible oil.edible oil.

edible oil.

• Detrimental Detrimental Detrimental

Detrimental

impact on impact on impact on

impact on

food food food

food

industries industries industries

industries

could be could be could be

could be

solved.solved.solved.

solved.

How much How much How much

How much

are PEs present in J. curcas seed oil?are PEs present in J. curcas seed oil?are PEs present in J. curcas seed oil?

are PEs present in J. curcas seed oil?

The amounts of PEs present in J. curcas seed oil vary from its source (approx. 0.01-1.58%).

SourcesSourcesSources

Sources

Phorbol esters , % (w/w)Phorbol esters , % (w/w)Phorbol esters , % (w/w)

Phorbol esters , % (w/w)

ReferencesReferencesReferences

References

Malaysia 0.23 W. A. Ahmed et al. , Phorbol Ester as Toxic Constituents of Tropical Jatropha Curcas Seed Oil, European Journal of

Scientific Research ISSN 1450-216X Vol.31 No.3 (2009), pp.429-436

Indonesia 1.58

India 0.58

Vietnam

Son La 0.62

Research Report for NEDO’s Northern part 0.19

Binh Thuan 0.27

Thailand

Chiang Mai 0.01Donlaporn Saetae and Worapot Suntornsuk, Variantion of Phorbol Ester Contents in Jatropha curcas from Different Provinces in Thailand and the Application of its Seed Cake

for Starter Broiler Diets, American-Eurasian J. Agric. & Environ. Sci., 8 (5): 497-501, 2010

Phrae 0.03

Phitsanulok 0.02

Satun 0.03

Oil Refining: Oil Refining: Oil Refining:

Oil Refining:

Role and Reduction/degradation of PEsRole and Reduction/degradation of PEsRole and Reduction/degradation of PEs

Role and Reduction/degradation of PEs


Sources:

H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of

H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of

Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173

Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173

–––

–

181181181

181

Oil RefiningOil RefiningOil Refining

Oil Refining

DegummingDegummingDegumming

Degumming

DeacidificationDeacidificationDeacidification

Deacidification

BleachingBleachingBleaching

Bleaching

DeodorizationDeodorizationDeodorization

Deodorization

Refined Refined Refined

Refined

OilOilOil

Oil

Degummed oilDegummed oilDegummed oil

Degummed oil

Acid gums Acid gums Acid gums

Acid gums

and and and

and

waste waterwaste waterwaste water

waste water

Distillation at 260oC under vacuum to remove undesirable volatile and odoriferous materials.

Interacting with bleaching agents to remove undesirable coloured impurities, remaining trace phospholipids, soap, metals and oxidation products.

Reacting with NaOH to neutralize remaining H3PO4 and/or Free fatty acids.

Interacting with H3PO4 to remove phosphatides in the form of gum.

ParametersParametersParameters

Parameters

Solvent Solvent Solvent

Solvent

extractedextractedextracted

extracted

PressedPressedPressed

Pressed

Silica-treated oilSilica-treated oilSilica-treated oil

Silica-treated oil

2.51 2.51 2.51

2.51

±±±

±

0.33 0.33 0.33

0.33

3.76 3.76 3.76

3.76

±±±

±

0.50 0.50 0.50

0.50


Parameters

SolventSolventSolvent

Solvent

extracted extracted extracted

extracted


Pressed

Crude oilCrude oilCrude oil

Crude oil

3.10 3.10 3.10

3.10

±±±

±

0.25 0.25 0.25

0.25

3.77 3.77 3.77

3.77

±±±

±

0.03 0.03 0.03

0.03


Parameters

Solvent extractedSolvent extractedSolvent extracted

Solvent extracted


Pressed

Acid gumsAcid gumsAcid gums

Acid gums

2.02 2.02 2.02

2.02

±±±

±

0.07 0.07 0.07

0.07

3.35 3.35 3.35

3.35

±±±

±

0.00 0.00 0.00

0.00

Wash waterWash waterWash water

Wash water

2.72 2.72 2.72

2.72

±±±

±

0.01 0.01 0.01

0.01

2.08 2.08 2.08

2.08

±±±

±

0.48 0.48 0.48

0.48


Parameters


Solvent


extracted


Pressed

Degummed oil 2.48 2.48 2.48

2.48

±±±

±

0.24 0.24 0.24

0.24

3.62 3.62 3.62

3.62

±±±

±

0.19 0.19 0.19

0.19


Parameters


Solvent


extracted


Pressed

Stripped oil NDNDND

ND

NDNDND

ND

Are Are Are

Are

there any risks of J. curcas PEs to environment?there any risks of J. curcas PEs to environment?there any risks of J. curcas PEs to environment?

there any risks of J. curcas PEs to environment?


Sources:

H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of

H. Makkar et al., Removal and Degradation of Phorbol Esters during Pre-treatment and Transesterification of

Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173

Jatropha curcas Oil, J Am Oil Chem Soc (2009) 86:173

–––

–

181181181

181

J. J. J.

J.

Curcas Curcas Curcas

Curcas

seedseedseed

seed

Oil Oil Oil

Oil

extractioextractioextractio

extractio

nnn

n

Crude Crude Crude

Crude

OilOilOil

Oil

Oil Oil Oil

Oil

RefiningRefiningRefining

Refining

FertilizersFertilizersFertilizers

Fertilizers

Live-stock Live-stock Live-stock

Live-stock


feeding

DegummingDegummingDegumming

Degumming

Degummed oilDegummed oilDegummed oil

Degummed oil

BleachingBleachingBleaching

Bleaching

DeodorizationDeodorizationDeodorization

Deodorization

Refined OilRefined OilRefined Oil

Refined Oil

TransesterificationTransesterificationTransesterification

Transesterification

Crude Crude Crude

Crude

BiodieselBiodieselBiodiesel

Biodiesel

Glycerol & ImpuritiesGlycerol & ImpuritiesGlycerol & Impurities

Glycerol & Impurities

Washing Washing Washing

Washing

with waterwith waterwith water

with water

Waste Waste Waste

Waste

waterwaterwater

water

Clean Clean Clean

Clean

BiodieselBiodieselBiodiesel

Biodiesel

DeacidificationDeacidificationDeacidification

Deacidification

Acid gums and wastewaterAcid gums and wastewaterAcid gums and wastewater

Acid gums and wastewater

Seed Seed Seed

Seed

cakecakecake

cake

• The presence of phorbol esters in the acid gums renders this fraction unsuitable for use in animal feed.

• The washings obtained during the degumming process are rich in phorbol ester and their disposal into the environment needs due care.

• the risk of PE for the people who have to work with these compounds or for humans who come into contact with treated water

Degradation of PEs in wastewater from degumming Degradation of PEs in wastewater from degumming Degradation of PEs in wastewater from degumming

Degradation of PEs in wastewater from degumming

processprocessprocess

process

Phorbol esters (PEs): Phorbol esters (PEs): Phorbol esters (PEs):

Phorbol esters (PEs):

Molecular StructureMolecular StructureMolecular Structure

Molecular Structure



20

TetraDecanoyl Phorbol-13-Acetae (TPA)TetraDecanoyl Phorbol-13-Acetae (TPA)TetraDecanoyl Phorbol-13-Acetae (TPA)

TetraDecanoyl Phorbol-13-Acetae (TPA)

OC

CH2OHH

OH

H3C

O

H

OH

H3C

CH3CH3

H

CCH3

H3C(H2C)11H2CC

O

O

O

• The PEs’ structure is dependent on the tetracyclic diterpene carbon skeleton (Tigliane).• PEs are hydrophobic, oil soluble and head stable when present in oil or seed cake.

A

B

C

D

1

23

4 56

78

910

11

1213

1415

1617

18

19

20

TiglianeTiglianeTigliane

Tigliane

HOHO

CH2OH

H

OH

H3C

O

H

OH

H3C

CH3CH3

phorbol

H


Phorbol

21

PEs: PEs: PEs:

PEs:

PEs: PEs: PEs:

PEs:

Degradation aspectsDegradation aspectsDegradation aspects

Degradation aspects


Degradation aspects

Biodiesel Biodiesel Biodiesel

Biodiesel

productioproductioproductio

productio

nnn

n

Crude J. curcas OilCrude J. curcas OilCrude J. curcas Oil

Crude J. curcas Oil

70-75% PEs70-75% PEs70-75% PEs

70-75% PEs

Free Fatty Free Fatty Free Fatty

Free Fatty

Acid Methyl Acid Methyl Acid Methyl

Acid Methyl

EstersEstersEsters

Esters

Glycerin, Glycerin, Glycerin,

Glycerin,


Phorbol

and and and

and

etc.etc.etc.

etc.

Transesterification Transesterification Transesterification

Transesterification

MeOH/AlkalineMeOH/AlkalineMeOH/Alkaline

MeOH/Alkaline

During Transesterification of J. curcas oil containing PEs, PEs are During Transesterification of J. curcas oil containing PEs, PEs are During Transesterification of J. curcas oil containing PEs, PEs are

During Transesterification of J. curcas oil containing PEs, PEs are

converted into Phorbol dissolving into glycerin phase.converted into Phorbol dissolving into glycerin phase.converted into Phorbol dissolving into glycerin phase.

converted into Phorbol dissolving into glycerin phase.

OC

CH2OH

H

OH

H3C

O

H

OH

H3C

CH3CH3

H

CCH3

H3C(H2C)11H2CC

O

O

O

HOHO

CH2OH

H

OH

H3C

O

H

OH

H3C

CH3CH3

phorbol

H

22

PEs: PEs: PEs:

PEs:

PEs: PEs: PEs:

PEs:


Degradation aspects


Degradation aspects

PEs containing in J. curcas seed cake can possibly degraded by PEs containing in J. curcas seed cake can possibly degraded by PEs containing in J. curcas seed cake can possibly degraded by

PEs containing in J. curcas seed cake can possibly degraded by

several means proposed by some authors. Then it can be used for several means proposed by some authors. Then it can be used for several means proposed by some authors. Then it can be used for

several means proposed by some authors. Then it can be used for

livestock feed.livestock feed.livestock feed.

livestock feed.

Livestock Livestock Livestock

Livestock


feeding

J. Curcas seed cakeJ. Curcas seed cakeJ. Curcas seed cake

J. Curcas seed cake

20-25% PEs20-25% PEs20-25% PEs

20-25% PEs

Degradation by some approachDegradation by some approachDegradation by some approach

Degradation by some approach

Treatment methodTreatment methodTreatment method

Treatment method

ConditionConditionCondition

Condition

(%) Removal or(%) Removal or(%) Removal or

(%) Removal or

DegradationDegradationDegradation

Degradation


References

stripping/deodorization at 260oC, 3 mbar and 1% steam injection

~ 100 Makkar et al. (2009)

alkali and heat treatment [1:1 (w/v) alkali, autoclaving at 121oC].

89 Rakshit et al. (2008)

Extraction with methanol and treatment with 0.07% NaHCO3

97 Martınez-Herrera et al. (2006).

Washing with methanol Autoclave heating at 121oC 95 Aregheore et al. (2003)

Solid State Fermentation (SSF) Pseudomonas aeruginosa PseA strain, 9 days

~ 100 Chetna Joshi et al. (2011)

Air bubbling autoclaving before and after treatment.

78.53 Sh. A. El Rafei et al (2011)

NaHCO3 and ozonation Ozone does: 50 mg/L, 3 min. 75.26 Sh. A. El Rafei et al (2011)

Γ-irradiation γ-ray dose of 50 kGy for 30 min. 71.35 Sh. A. El Rafei et al (2011)

What What What

What

are the new methods proposed to do?are the new methods proposed to do?are the new methods proposed to do?

are the new methods proposed to do?

According to literature reviewAccording to literature reviewAccording to literature review

According to literature review

, the degradations of PEs the degradations of PEs the degradations of PEs

the degradations of PEs

are based on the conventional and aaa

a

dvance ooo

o

xidative processesprocessesprocesses

processes

(AOP) such as

• combination with chemical and heat treatmentcombination with chemical and heat treatmentcombination with chemical and heat treatment

combination with chemical and heat treatment

, , ,

,

• biological oxidation using microbesbiological oxidation using microbesbiological oxidation using microbes

biological oxidation using microbes

, , ,

,

• the combination of Aeration and heat treatmentthe combination of Aeration and heat treatmentthe combination of Aeration and heat treatment

the combination of Aeration and heat treatment

, , ,

,

• the combination of chemical and Ozonation the combination of chemical and Ozonation the combination of chemical and Ozonation

the combination of chemical and Ozonation

or or or

or

-irradiation-irradiation-irradiation

-irradiation

...

.

New methods are based on AOPNew methods are based on AOPNew methods are based on AOP

New methods are based on AOP

• Ultrasound irradiation: Sonolytic degradationUltrasound irradiation: Sonolytic degradationUltrasound irradiation: Sonolytic degradation

Ultrasound irradiation: Sonolytic degradation

• Cold Plasma irradiationCold Plasma irradiationCold Plasma irradiation

Cold Plasma irradiation

•

-ray irradiation: -ray irradiation: -ray irradiation:

-ray irradiation:

-Radiolytic degradation of -Radiolytic degradation of -Radiolytic degradation of

-Radiolytic degradation of

PEs.PEs.PEs.

PEs.

• The first two methods have not been studies by any author on the degradation of Jatropha PEs.• Even though -irradiation method has been used for degradation of PEs in the J. curcas seed cake, there is

not any work on the degradation of PEs in liquid media such as in oil or other solvents as well as in water.

L. WojnL. WojnL. Wojn

L. Wojn

ááá

á

rovits, E. Takrovits, E. Takrovits, E. Tak

rovits, E. Tak

ááá

á

cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry

cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry

77 (2008) 22577 (2008) 22577 (2008) 225

77 (2008) 225

–––

–

244244244

244

Advance Oxidation Processes (AOP)Advance Oxidation Processes (AOP)Advance Oxidation Processes (AOP)

Advance Oxidation Processes (AOP)

L. WojnL. WojnL. Wojn

L. Wojn

ááá

á

rovits, E. Takrovits, E. Takrovits, E. Tak

rovits, E. Tak

ááá

á

cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry

cs, Irradiation treatment of azo dye containing wastewater: An overview,Radiation Physics and Chemistry

77 (2008) 22577 (2008) 22577 (2008) 225

77 (2008) 225

–––

–

244244244

244

Reaction Reaction Reaction

Reaction

Effect ofEffect ofEffect of

Effect of

Saturation Saturation Saturation

Saturation

AtmosphereAtmosphereAtmosphere

Atmosphere

pH pH pH

pH

RangeRangeRange

Range

AdditivesAdditivesAdditives

Additives

Related reactionsRelated reactionsRelated reactions

Related reactions

•OH, H• and eaq

N2 or Ar 3 - 11 - H2O → eaq ; •OH; •H

•OH, H• N2 or Ar < 2 - eaq + H3O+ → H• + H2O

H• N2 or Ar < 2 t-butanol0.2–1 mol/dm-3

slow: H• +(CH3)3COH → •CH2(CH3)2COH + H2

•OH + (CH3)3COH → •CH2( CH3)2COH + H2O

eaq N2 or Ar > 3 t-butanol0.2–1 mol/dm-3

fast: H• +(CH3)3COH → •CH2(CH3)2COH + H2

•OH + (CH3)3COH → •CH2( CH3)2COH + H2O

•OH N2O 3 - 11 - eaq + N2O + H2O → •OH + OH + N2

•OH, O2•/HO2•

Air or O2 - -eaq + O2 → O2•

H• + O2 → HO2•O2• + H3O + ↔ 2HO2• + H2O

O2•/HO2• O2 - t-butanol0.2–1 mol/dm-3

-

•CH2(CH3)2COH N2O > 3 t-butanol0.2–1 mol/dm-3

-

Free Radical Effect DescriptionFree Radical Effect DescriptionFree Radical Effect Description

Free Radical Effect Description

No.No.No.

No.

Treatment MethodsTreatment MethodsTreatment Methods

Treatment Methods

Removal/degRemoval/degRemoval/deg

Removal/deg

radation (%)radation (%)radation (%)

radation (%)


References

1. 121oC, 30 min → 4 times washing with MeOH. 95 E. M. Aregheore et al., Detoxification of a toxic variety of Jatropha curcas using heat and chemical treatments, and preliminary nutritional evaluation with rats, S. Pac. J. Nat. Sci., 2003, 21, 50-56

2. 4% NaOH + 10% NaOCl → 121oC, 30 min. 93

3. 3.5% NaOH → 121oC, 30 min. 90

4. 0.07% NaHCO3 → 121oC, 25min. 75 J. Martínez-Herrera et al., Chemical composition, toxic/ antimetabolic constituents, and effects of different treatments on their levels, in four provenances of Jatropha curcas L. from Mexico, Food Chemistry 96 (2006) 80–89.

5. 0.07% NaHCO3 → -irradiation, 10 kGy 18

6. Extraction with EtOH, room temperature, 2h. 96

7. 2% NaOH/Ca(OH)2 → 121oC, 30 min. → water dispersion, 1h. 88-90

K.D. Rakshit et al., Toxicity studies of detoxified Jatropha meal (Jatropha curcas) in rats ,Food and Chemical Toxicology 46 (2008) 3621–3625.

8. Enzymatic oxidation in soil, 21 days. 100Rakshit K. D. et al., Biodegradation of Jatropha curcas phorbol esters in soil, J Sci Food Agric 2010; 90: 2090–2097

10. Solid-state fermentation using Pseudomonas aeruginosa PseA strain, 9 days. 100

C. Joshi et al., Degradation of phorbol esters by Pseudomonas aeruginosa PseA during solid-state fermentation of deoiled Jatropha curcas seed cake, Bioresource Technology 102 (2011) 4815–4819.

11. 0.2 N NaHCO3 → Ozone dose 50 mg/L, 2-3 min. 75.26Sh. A. El Rafei et al., Ozone for Phorbol Esters Removal from Egyptian Jatropha Oil Seed Cake, Adv. Appl. Sci. Res., 2011, 2 (4):221-232

12. 121oC, 30 min → Aeration, 3 min. → 121oC, 25 min. 78.53

13. 0.2 N NaHCO3 →

-irradiation, 50 kGy.-irradiation, 50 kGy.-irradiation, 50 kGy.

-irradiation, 50 kGy.

71.35

Are Are Are

Are

there any published methods of degrading J. curcas PEs?there any published methods of degrading J. curcas PEs?there any published methods of degrading J. curcas PEs?

there any published methods of degrading J. curcas PEs?

Phorbol esters: Phorbol esters: Phorbol esters:

Phorbol esters:

Proposed Treatment Methods : Proposed Treatment Methods : Proposed Treatment Methods :

Proposed Treatment Methods :

Ultrasonic irradiation Ultrasonic irradiation Ultrasonic irradiation

Ultrasonic irradiation

and and and

and

γ-ray radiation-ray radiation-ray radiation

-ray radiation



27

Basic PrincipleBasic PrincipleBasic Principle

Basic Principle

Ultrasonic irradiation (US)Ultrasonic irradiation (US)Ultrasonic irradiation (US)

Ultrasonic irradiation (US)

γ-ray radiation (-ray radiation (-ray radiation (

-ray radiation (

γ-ray)-ray)-ray)

-ray)

• produces strong cavitation in aqueous solution:• shock wave. • reactive free radicals (e.g., •OH, HO2•, and O•) by violent collapse of the cavitation bubble.• formation of H2O2.

• in aqueous solution, it generates: • hydrated electrons, hydrogen atoms, and • hydroxyl radicals as initial radical species.• hydrogen and hydrogen peroxide (H2O2) are

formed by their recombination reactions.

The decomposition of toxic chemicals should be contributed by these effects.

Study on Degradation of Phorbol estersStudy on Degradation of Phorbol estersStudy on Degradation of Phorbol esters

Study on Degradation of Phorbol esters

200 kHz Sonicator 60Co as γ-ray source

At different powers, amplitudes, times At different doses, times

HPLC/UV-Vis analysis of phorbol ester concentration before and after irradiation

HPLC/UV-Vis analysis of phorbol ester concentration before and after irradiation

LC-MS analysis for confirmation of compounds LC-MS analysis for confirmation of compounds

Colorimetric analysis of H2O2 formation Colorimetric analysis of H2O2 formation



28

The similar bonding structure shows their similarity in The similar bonding structure shows their similarity in The similar bonding structure shows their similarity in

The similar bonding structure shows their similarity in

reactionreactionreaction

reaction

OC

CH2OH

H

OH

H3C

O

H

OH

H3C

CH3CH3

H

CCH3

H3C(H2C)11H2CC

O

O

OCH2

CH

H2C OCR3

O

R1OCO

R2OCO

(TPA or PMA)(TPA or PMA)(TPA or PMA)

(TPA or PMA)

(Triglycerides)(Triglycerides)(Triglycerides)

(Triglycerides)



29

Simplification of Phorbol Ester StructureSimplification of Phorbol Ester StructureSimplification of Phorbol Ester Structure

Simplification of Phorbol Ester Structure

OC

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

CCH3

H3C(H2C)11H2CC

O

O

O

RRR

R

111

1

RRR

R

222

2

OC

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

C

R1

O

O

O

R2

Extraction of J. curcas phorbol esterExtraction of J. curcas phorbol ester• 10 g of Jatropha oil was

extracted in triplicates each with 10 mL of methanol to give 3 fractional extracts.



• 2 mL of each fraction was kept for analyzing fractional extraction profile.

• The rest of each fraction was combined.


Fig. 1

illustration of the procedure on the extraction of phorbol esters from Jatropha oil.

FractionExtraction

(%)Conc. (ppm)

1st 57 1994.962nd 29 999.903rd 14 478.48


Fig. 6

HPLC/UV chromatogram at 280 nm of each methanolic fractions containing phorbol esters obtained from extraction of Jatropha oil.

• Only one extraction is not enough to get J. curcas phorbol esters from the J. curcas oil.

• There were 2 groups of peaks appear at the retention time: the 1st group (unknown) was between 5.5 and 7 min; and the 2nd group (Jatropha phorbol esters) between 8.5 and 9.5 min.



Transesterification of J. curcas phorbol esterTransesterification of J. curcas phorbol ester• 2 mL of methanolic fraction containing J. curcas PEs was added onto a graduated

tube.• 0.4 mL of 10%(w/w) KOH in methanol was further added the methanolic sample in

the tube.• The mixture was gently shaken for 1 min to get homogeneous mixture. Then it was

kept in a dark place for 14 hours to complete reaction.• After completing reaction, the mixture was neutralized with 85%

H3PO4 to the pH ~ 7.

• The light-yellowish solution was transferred onto a vial for

analysis. • The light-yellowish solution was

filtered with filter-unit before

analysis by HPLC/PDA.

• After analysis, white precipitate

was found at the bottom of the vial.




Fig. 4

Trans-esterifying methanolic extract containing Jatropha phorbol esters.


Fig. 7

HPLC/UV chromatogram at 280nm of methanolic fraction before and after transesterification.

• Before transesterification the Jatropha phorbol ester appears at retention time between 8-10 min.

• After transesterification the peaks of Jatropha phorbol esters disappear due to conversion into FAME.

• The concentration of J. curcas PEs was 3.47 0.07 mg-PEs/g-Oil.





36

KOH + CH3OH CH3O- + KH+

OC

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

C

R1

O

O

O

R2

CH3O-

CH3O-

OC

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

C

R1

O-

O-

O

R2

CH3O

CH3O

OC

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

C

R1

O-

O-

O

R2

CH3O

CH3O

-O

C

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

C

R1

O

O

-O

R2

CH3O

CH3O

-O

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

-O

KH+

KH+

HO

CH2OHH

OH

H3CO

H

OH

H3C

CH3CH3

H

HO

K*

K*+

Proposed Mechanism on the conversion of phorbol ester to phorbol Proposed Mechanism on the conversion of phorbol ester to phorbol Proposed Mechanism on the conversion of phorbol ester to phorbol

Proposed Mechanism on the conversion of phorbol ester to phorbol

during transesterification of J. oil in Biodiesel productionduring transesterification of J. oil in Biodiesel productionduring transesterification of J. oil in Biodiesel production

during transesterification of J. oil in Biodiesel production

Sonolytic Degradation of Jatropha PEsSonolytic Degradation of Jatropha PEsSonolytic Degradation of Jatropha PEs

Sonolytic Degradation of Jatropha PEs

Sonolysis system for chemical degradation study: (a) front Sonolysis system for chemical degradation study: (a) front Sonolysis system for chemical degradation study: (a) front

Sonolysis system for chemical degradation study: (a) front

side; (b) top side; (c) Model and max frequency; (d) side; (b) top side; (c) Model and max frequency; (d) side; (b) top side; (c) Model and max frequency; (d)

side; (b) top side; (c) Model and max frequency; (d)

Temperature control.Temperature control.Temperature control.

Temperature control.

Scope of ResultsScope of ResultsScope of Results

Scope of Results

•Degradation at Xn mg/L and kinetic data

(ln(C0/C).

•UV adsorption spectra before and after

degradation

•Degradation vs. power density.

•Effect of pH, temperature, additives.

•Ratio of H2O2 to PEs on degradation.

•etc.

(a)(a)(a)

(a)

(b)(b)(b)

(b)

(d)(d)(d)

(d)

(c)(c)(c)

(c)

Radiolytic Degradation of Jatropha PEsRadiolytic Degradation of Jatropha PEsRadiolytic Degradation of Jatropha PEs

Radiolytic Degradation of Jatropha PEs

Radiolytic Radiolytic Radiolytic

Radiolytic

Sampling racksSampling racksSampling racks

Sampling racks

A water-pool A water-pool A water-pool

A water-pool

606060

60

Co Co Co

Co

-ray irradiation facilities at -ray irradiation facilities at -ray irradiation facilities at

-ray irradiation facilities at

the Radiation Research Center, Osaka the Radiation Research Center, Osaka the Radiation Research Center, Osaka

the Radiation Research Center, Osaka

Prefecture University, Japan.Prefecture University, Japan.Prefecture University, Japan.

Prefecture University, Japan.

Cherenkov radiation Cherenkov radiation Cherenkov radiation

Cherenkov radiation

from the from the from the

from the

606060

60

Co g-ray Co g-ray Co g-ray

Co g-ray

sourcesourcesource

source

Scope of ResultsScope of ResultsScope of Results

Scope of Results

•Absorbed Dose: Absorbed Dose: Absorbed Dose:

Absorbed Dose:

degradation G-value and constant.•UV absorption: UV absorption: UV absorption:

UV absorption:

degradation spectra on UV adsorption•Effect of additives and saturation atmosphere vs. Effect of additives and saturation atmosphere vs. Effect of additives and saturation atmosphere vs.

Effect of additives and saturation atmosphere vs.

degradationdegradationdegradation

degradation

•etc.etc.etc.

etc.




Fig. 9

HPLC/UV chromatogram of TPA’s 30ppm (a-b) and 50ppm (c-d) at r-ray absorption dose of 0, 10 and 20kGy, respectively.

(((

(

a)a)a)

a)

(((

(

b)b)b)

b)

(((

(

c)c)c)

c)

(((

(

d)d)d)

d)

• After exposure to -irradiation, the TPA was degraded.

• From HPLC/UV chromatogram at 220nm around 50% TPA was degraded at 10 kGy. At 20kGy, TPA degradation was greater than 90% .

• In contrast, 56% TPA was observed at HPLC/UV at 280nm when the TPA was irradiated at 10 kGy and complete degradation was observed at 20 kGy -irradiation.

-Radiolytic Degradation of TPA-Radiolytic Degradation of TPA• 30 and 50 ppm-TPA solutions was prepared by using 1000ppm TPA standard

solution dissolved in methanol.• Vials each separately containing methanol, 30-and-50ppm TPA solution was

divided into two parts for 10 and 20 kGy, respectively.

• The vials were put onto the stainless steel sample holder and exposed to -ray at absorbed dose of 10 and 20kGy, respectively.

• The vial’s color was changed to brown after r-ray exposure.




Fig. 4

-irradiation of TPA solution at irradiation rate of ~ 10 kGy/h.

Results on Degradation of Jatropha PEsResults on Degradation of Jatropha PEsResults on Degradation of Jatropha PEs

Results on Degradation of Jatropha PEs

Effect of Effect of Effect of

Effect of

-Irradiation-Irradiation-Irradiation

-Irradiation

Documents

(201 3/2/1 Nagoya) - 大阪府立大学matsu/data/isplasma5/...3 Phorbol Esters Phorbol Esters Introduction Introduction Introduction Introduction J. Curcas J. Curcas seed seed kernel