Profiling Biomass Resources And Their Chemistriesbiorefinery.utk.edu/posters/Biomass...

Profiling Biomass Resources And Their Chemistries

Art J. RagauskasSchool of Chemistry and Biochemistry

Georgia Institute of Technology


Global Biomass Opportunities220 billion tons of dry biomass available

0.4% of all biomass is for food


USA Biomass Reserves

SyngasFischer Tropsch FuelBio-EthanolBio-ButanolDMEBiodiesel/BiogasolinePyrolysis Oils

Profiling Biomass Resources: Biopolymers

OO

HOOH

OO

HOOH

OH

OH

O

HOOH

OO

HOOH

O

OH

OH

O

10,000 - 750

OO

O OAcO

O

HOOAc

O

HOO

OO

AcOOH

O-XylanO

O

HOOAc

OO

AcOOAc

O

O

HO

HO

OHO

OHO

HO OH

HO

O

O

Ph

OH

O

O

O

H3CO

OH

OH

OH

DP: 70 - 200

Cellulose is the most abundant organic chemical on earth, Annual biosphere production of ≈90x109 metric tons.

HemicellulosesMain Sugars: Xylose, Glucose, Glucouronic acid, Arabinose, Galactose, Mannose

OH

OH

R R'

Enzymatic Polymerization for SW

R,R' Alcohol Name Source

H,H p-coumaryl Compression Wood, Grasses

H, OCH3 coniferyl Softwood and Hardwood

OCH3, OCH3 sinapyl Hardwoods

Lignin

DPWood:60 -100

Profiling Biomass Resources: Minor Components

CO2H CO2H

Pimaric Abietic

CO2H

Palmitic Acid

CO2H

Oleic Acid

• From Wood to Fuels: Integrating biofuels and pulp production• Ragauskas, A.J., Nagy, M.; Kim, D.H.; Eckert, C.A.; Liotta, C.L. Industrial Biotechn., (2006), 2(1), 55-65.

• Profiling the wood and pulping properties of southern pine thinning resources. • Ragauskas, A.J.; Sealey, J. et al TAPPI Journal (2005), (1), 21-25.

• Current and Future Softwood Kraft Lignin Process Chemistry. • Chakar, F.S.; Ragauskas, A.J. Industrial Crops and Products (2004), 20(2), 131-141.

• Intrinsic Metal Binding Capacity of Kraft Lignins. • Werner, J.; Ragauskas, A.J.; Jiang, J.E., Journal of Wood Chem. Techn. (2000), 20(2), 133-145.

• Metal Profiling of Southeastern U.S. Softwood and Hardwood Furnish. • Allison, L.; Ragauskas, A.J.; Hsieh, J., TAPPI Journal (2000), 83(8), 97-102.

HO

Sitosterol

Pine

Wood Pine

Profiling Biomass ChemistriesPlant Carbohydrates - Structure

Profiling Biomass ChemistriesCellulosic FibersLignocellulosic Fibers

SEM-TEM-AFM

Profiling Biomass Resources and Chemistries

Cellulose


CelluloseI – IV PolymorphsAmphorousParacrystalline

Cellulose Iα and Iβ

Chains are parallelDifferences due to Orientation of

Cellulose Sheets

Cellulose II:

antiparrellelNishiyama, Y., et al Journal of the American

Chemical Society (2003), 125(47), 14300


Cellulose: Iα is more abundant in lower plants/bacteriaIβ is more abundant in higher plants

Analysis of Cellulose Crystallinity: X-ray, NMR, FT-IR

O

OO

CH2OH

OHO

OH

HOOH

CH2OH1

2

3

45

6

4

1

C-1

C-4

C-2, 3, 5

C-6

Assignments Chemical shift (ppm)

FWHH* (Hz)

Cellulose I(α) 89.5 24 Cellulose I(α+β) 88.8 198 Para-crystalline cellulose 88.3 234 Cellulose I (β) 87.8 26 Accessible fibril surface 84.3 64 Inaccessible fibril surfaces and hemicellulose 83.7 467

Accessible fibril surface 83.3 0.8


Cellulase Treatment

0 5 10 15 20 253

4

5

6

7

8

Rel

ativ

e in

tens

ity, %

Hydrolysis time, hr

ΙβCellulose: Iβ

0

20

40

60

80

100

0 10 20 30 40 50

Hydrolysis Time, h

Glu

cose

yie

ld, %

Pulpavicel

800 1000 1200 1400 1600 1800 2000

m/z

MALDI mass spectrum of Cellulase hydrolysis effluent consists of series of oligosaccharidescontaining 6,7,8,9,10 glucose.

Cellulose: Iα

0 5 10 15 20 252.5

3.0

3.5

4.0

4.5

5.0

Rel

ativ

e in

tens

ity, %

Hydrolysis time, hr

ΙαCellulose: Iα


0 5 10 15 20 2550

51

52

53

54

55

56

CrI

, %Hydrolysis time, hr

0

20

40

60

80

100

0 10 20 30 40 50

Hydrolysis Time, h

Post

-hyd

roly

sis p

erce

ntag

e, %

avicelpulp

0 5 10 15 20 25

33

34

35

36

37

Rel

ativ

e in

tens

ity, %

H yd ro lysis tim e, h r

P ara-cry sta lline

0 5 10 15 20 25

46

48

50

Rel

ativ

e in

tens

ity, %

Hydrolysis time, hr

Amorphous

CP/MAS 13C NMR analysis of cellulase treated bleached softwood kraft pulp. Ragauskas, et al.. Carbohydrate Research (2006), 341(5), 591-597.


Lignin


CH

OH

OCH3

CH

CH2OH

GUAIACYL

CH

OH

OCH3

CH

CH2OH

CH3O

CH

OH

CH

CH2OH

SYRINGYL p-Coumaryl alcohol

12

34

5

6

γ

β

α

CH

CH

CH2OH

OCH3

O

OCH3

O

+

OCH3

O

HCOR

HC

HOH2C

OCH3

OH1

2

4

OCH3

O

HCOR

HC

HOH2C

OCH3

O

OCH3

O

HCOR

HC

HOH2C

OCH3

OH

CH

CH

CH2OH

OCH3

O

+

5 6 1

3

ROH

OCH3

O

CH

HC

HOH2C

OCH3

O

HC

HC

HOH2C

OCH3

O

OCH3

O

HCOR

HC

HOH2C

OCH3

O

H

HC

HC

HOH2C

OCH3

OH

OCH3

O

HCOR

HC

HOH2C

OCH3

O

7 8


MW: Lignin samples are acetylated dissolved in tetrahydrofuran and GPC analyzed using HP 1090 Liquid chromatography containing ultraviolet diode array detector and HP 1047A refractive index detector.

Starting: ∼ 11,000


Corn stover

Switchgrass

• Ball Milling• Acidic Dioxane Solvent Lignin• Cellulase Purification


% SW HW

A: ß-O4 50 60

B: α-O4 8 ta

C: 15 5

D: ß-5 11 6

E: 5-5 10 5

F: 5-O4 4 7

G: ß-1 7 7

H: ß- ß 2 3

C

C O

C

C

C

C

O

CH3O

OCH3O OCH3

O

CH2OH

O

CH3O

C

C

C

OO

CH3O OCH3O

OCH3O O

OCH3

C

C

C

O

OCH3

CH3O

O

O

A B C

D

E

F

G

H


φφ

OCHOCH33

φφ--OHOH

C=OC=ORORROR

DMSODMSO

φφ--CHCH22--φφ

180 160 140 120 100 80 60 40 PPM

Pine 13C CP/MAS NMR


O

C OH

CH

OCH3

CH2OH

H

CH3OO

OCH3O

H

O

C O-CH

OCH3

CH2OH HO-

O

C

C

OCH3

CH2OH

OH

CH3OO-

H

+

Alkaline Depolymerization of Lignin


φφ

OCHOCH33

φφ--OHOH

C=OC=ORORROR

DMSODMSO

φφ--CHCH22--φφ

φφ

OCHOCH33

φφ--OHOH

C=OC=ORORROR

DMSODMSO

φφ--CHCH22--φφConjugatedcarbonyl

20406080100120140160180 0ppm

Saturatedcarbonyl

Aromatic and Olefinic C

Aliphatic C-O

Muconic acidOCH3

D residual lignin

DE effluent lignin

DE residual lignin

Aromatic OCH3Conjugatedcarbonyl

20406080100120140160180 0ppm

Saturatedcarbonyl

Aromatic and Olefinic C

Aliphatic C-O

Muconic acidOCH3

D residual lignin

DE effluent lignin

DE residual lignin

Aromatic OCH3


ppm024 6 8 10 12

DED residual lignin

1H NMR


150 148 146 144 142 140 138 136 ppm

OMeOH

OMeOH

HO R

OH

R

O

OH

CyclohexanolCyclohexanol: : Internal Internal standardstandard

150 148 146 144 142 140 138 136 ppm150 148 146 144 142 140 138 136 ppm

OMeOH

OMeOH

HO R

OH

R

O

OH

CyclohexanolCyclohexanol: : Internal Internal standardstandard

+O

O

CH3CH3

CH3

CH3P Cl Lignin-OH

OMeO

POO

O

O

CH3CH3

CH3

CH3P O Lignin

OMeO

POO

O

OP O R

O

ROPO

O

Kappa factor

-1.0-0.8-0.6-0.4-0.20.0

0 0.05 0.1 0.15 0.2

Guaiacyl phenolic

Condensed phenolic

Kappa factor

-1.0-0.8-0.6-0.4-0.20.0

0 0.05 0.1 0.15 0.2

Guaiacyl phenolic-1.0-0.8-0.6-0.4-0.20.0

0 0.05 0.1 0.15 0.2

Guaiacyl phenolic

Condensed phenolic

31P NMROf Lignin


O

O

P(OCH 3 )3O

O

P(OCH 3)3

OP

OOCH 3

OCH 3

OCH 3

I II III

Ramirez et al., Sidky et al., & MedveczRamirez et al., Sidky et al., & Medvecz

H 2 O

OR

OR

R = PO(OCH 3 )(OH) or H

O

O

P(OCH 3 )3O

O

P(OCH 3)3

OP

OOCH 3

OCH 3

OCH 3

I II III

O

O

P(OCH 3 )3O

O

P(OCH 3)3

OP

OOCH 3

OCH 3

OCH 3

I II III

Ramirez et al., Sidky et al., & MedveczRamirez et al., Sidky et al., & Medvecz

H 2 O

OR

OR


H 2 O

OR

OR


0 -5 -10 -15 -20 -25PPM

Quinone AdductQuinone Adduct

PO O

O

O

Internal StandardInternal Standard

0 -5 -10 -15 -20 -25PPM

Quinone AdductQuinone Adduct

PO O

O

OPO O

O

O

Internal StandardInternal Standard

0.00

0.05

0.10

0.15

0.20

0.25

Brownstock D D(EAr) DE D(EO) D(EP) D(EPO)

Bleach Stage

Qui

none

Con

tent

(mm

ol/g

isol

ated

lign

in)

Residual Effluent

O

O O

OOCH3

0.00

0.05

0.10

0.15

0.20

0.25

Brownstock D D(EAr) DE D(EO) D(EP) D(EPO)

Bleach Stage

Qui

none

Con

tent

(mm

ol/g

isol

ated

lign

in)

Residual Effluent

O

O O

OOCH3


-56 -57 -58 -59 -60 -61 -62 -63ppm

IS

Carbonyl

Quinone

+ H2NNH CF3O

CF3NNH

19F-NMR: Lignin

-56 -57 -58 -59 -60 -61 -62 -63ppm

IS

Carbonyl

Quinone

+ H2NNH CF3O

CF3NNH

19F-NMR: Lignin


230.0 300 350 400 450 500.0-0.10

0.0

0.1

0.2

0.3

0.4

0.50

NM

A

LAAOLAALAAOO7LAAOO3LAAOO9LAAOO1

OCH3

OH

HO

O

OH

OCH3

1. λmax 250 and 300 nm are assigned to unconjugated phenolics, decreases with aggressive O-stages conditions is consistent with the 31P NMR data.

2. Lignin samples appear to be relatively free of phenolic stilbenes (λmax375 nm).

3. λmax350 nm has been attributed to phenolic α-carbonyl groups

UV/Vis Ionization Difference Spectroscopy

230.0 300 350 400 450 500.0-0.10

0.0

0.1

0.2

0.3

0.4

0.50

NM

A

LAAOLAALAAOO7LAAOO3LAAOO9LAAOO1

OCH3

OH

HO

O

OH

OCH3

1. λmax 250 and 300 nm are assigned to unconjugated phenolics, decreases with aggressive O-stages conditions is consistent with the 31P NMR data.

2. Lignin samples appear to be relatively free of phenolic stilbenes (λmax375 nm).

3. λmax350 nm has been attributed to phenolic α-carbonyl groups

UV/Vis Ionization Difference Spectroscopy

FT-IR, Pyrolysis GC-MSNIR, TGA: Facile Identification

Other Techniques

1100.0 1500 2000 2500.00.00

0.1

0.2

0.3

0.4

0.50

NM

A

NIR Spectra of Lignocellulosics

• Structural Analysis of Acetylated Hardwood Lignin and Their Photoyellowing Properties.• Pu, Y.; Ragauskas, A.J. Canadian J. Chemistry. (2005), 83(12), 2132-2139.

• Biobleaching chemistry of laccase-mediator systems on high-lignin-content kraft pulps. • Chakar, F.S.; Ragauskas, A.J., Canadian Journal of Chemistry (2004), 82, 344-352.

• Oxygen Delignification Chemistry and Its Impact on Pulp Fibers.• Ragauskas, A.; Jameel, H., et ak Journal of Wood Chemistry Technology (2003), 23(1), 13-29.

• Comparative Evaluation of O-Delignification Processes for Low- High-Lignin-Content SW Kraft Pulps. • Ragauskas, A.J. et al, Industrial & Engineering Chemistry Research (2002), 41, 5171-5180.

• Oxygen Degradation and Spectroscopic Characterization of Hardwood Kraft Lignin. • Ragauskas, A. et al. Industrial & Engineering Chemistry Research (2002), 41(24), 5941-5948.


Hemicellulose


Wood HemicellulosesO

O

O OAcO

O

HOOH

O

HOO

OO

HOOH

O-XylanO

HOO

OOH

O

H3CO HO

OHHO2C O

HO

HOOH

OO

HOOH

OO

HO

OHO

OH

O

AcO

OH

OO

HO

OHOH

OH

O

O

HO HO

OH

HO

OO

HOOH

OO

HO

OHOH

OH

O

HOOH

OO

HO

OHOH

OH

O

Softwood - galactoglucomannan

Softwood - arabinoglucuronoxylan

Hardwood - glucomannan

Wood Species

Ara Xyl Gal Glc Man Rha GlcA GalA 4-O-MeGlcA

Totala %

Softwood Picea abies sapwood 0.14 0.61 0.17 0.37 1.00 0.02 0.03 0.16 0.10 24.6 heartwood 0.17 0.69 0.28 0.35 1.00 0.03 0.04 0.20 0.12 24.9 Pinus banksiana sapwood 0.18 0.57 0.18 0.40 1.00 0.02 0.05 0.13 0.10 27.2 heartwood 0.22 0.75 0.37 0.43 1.00 0.03 0.06 0.17 0.14 29.3 Hardwood Betula pendula stemwood 0.02 1.00 0.06 0.08 0.04 0.02 0.01 0.10 0.16 33.6% Populus tremuloides sapwood 0.03 1.00 0.04 0.11 0.05 0.03 0.01 0.12 0.13 29.1 heartwood 0.03 1.00 0.04 0.12 0.09 0.03 0.01 0.12 0.13 28.8


Xylan

10

30

50

70

90

110

0 2 4 6 8 10

Hydrolysis Time, h

Pos

t-hyd

roly

sis

perc

enta

ge, % Level 1(5000 u/g)

Level 2(20000 u/g)Level 3(50000 u/g)

OO

HOOR

OO

HOOR

O

HOOH

OO

HOOH

O-XylanO

n: 100 - 200

Native R Substituted with OH3CO

HOOH

O

H3CO2C

O OHO

HO

HO O

CH3Pulp R=H

SW Wood Pulp, Beech wood xylan

Xylanase pH 6.8, Rxn Temp. 38oC

0

10

20

30

40

50

0 2 4 6 8 10

Hydrolysis Time, h

Xyl

ose

yiel

d, %

xylan


10

30

50

70

90

110

0 2 4 6 8 10

Hydrolysis Time, h

Pos

t-hyd

roly

sis

perc

enta

ge, % Level 1(5000 u/g)

Level 2(20000 u/g)Level 3(50000 u/g)

MALDI spectrum of xylan after 8 hour hydrolysis by xylanase.

• Elucidating carboxylic acid profiles for extended oxygen delignification of high-kappa softwood kraft pulps. Zhang, D.; Pu, Y.; Chai, X. S.; Naithani, V.; Jameel, H.; Ragauskas, A.J. Holzforschung (2006), 60(2), 123-129.

• Influence of Hexenuronic Acids on U.S. Bleaching Operations.

Chakar, F.S.; Allison, L.; Ragauskas, A.J.; McDonough, T.J.; Sezgi, U., TAPPI Journal (2000), 83(11), 62-71.


Ragauskas Biomass Characterization

Topochemistry of Biopolymers: Staining/SEM, AFM

Cellulose: Challenge is to monitor crystallinity, DPHydrolyzed oligomers

Lignin: Need to determine structure/functionality/DP

Hemicellulose: Need to determine structure/functionality/DP

Extractives: Need chemical constituentsmetals, terpenes, lignans, fatty acids, etc

Biomass CharacterizationPath Forward

Biomass Characterization

Genetic Manipulation • Separations-Biomaterials• Liquidification• Biopower• Biofuels • SensorsRagauskas Biomass Expertise:

Macro & Chemical StructureDegree of Polymerization for Lignin/Hemicellulose/Cellulose

Analysis of metals/extractives

Thank You!

Profiling Biomass Resources And Their Chemistriesbiorefinery.utk.edu/posters/Biomass...

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