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Cellulose: structure, morphology, and

crystalline forms

CHEM-E2140

Eero Kontturi

29th October 2015


Outline
(1) Molecular structure of cellulose
- molecular vs. supramolecular structure

(2) Crystalline forms (allomorphs): - cellulose I


- cellulose II
- cellulose III
- cellulose IV
- consequences of the crystalline forms

(3) Morphology of native cellulose: microfibril


- models for microfibrillar cross sections
- crystalline-amorphous structure
- microfibril aggregation / coalescence
Molecular structure of cellulose
Non-reducing Reducing
end group end group
Reducing
OH OH end group
Anhydroglucose
O
OH O OH
O HO
HO HO O H
O O
HO HO n /2 OH
OH O OH
OH OH

Cellobiose

OH
• Reducing end group is in equilibrium HO OH
O
with a cyclic hemiacetal O

OH
Molecular vs. supramolecular structure
Molecular structure Supramolecular structure

OH
Anhydroglucose
OH O
HO O
O O
HO n /2
O OH
OH

Cellobiose

- (1→4)-β-D-glucopyranose - individual cellulose chains linked


- high native DP (~5000-15000) together by intermolecular bonding

Structure revealed: Structure revealed (cellulose Iα ja Iβ):


Freudenberg Nishiyama et al.
Liebigs Ann. Chem. 1928, 461, 130. J. Am. Chem. Soc. 2002, 124, 9074.
Haworth Nature 1930, 126, 438. J. Am. Chem. Soc. 2003, 125, 14300.
Crystalline forms of cellulose
Cellulose allomorphs
NATIVE
FORM Regeneration or
alkali treatment
Cellulose Iα
Cellulose II
Cellulose Iβ
IRREVERSIBLE

Liquid NH3 or REVERSIBLE


diamine treatment

Cellulose IIII Cellulose IIIII

Heat treatment

New evidence suggests that cellulose IV


Cellulose IVI is a slightly disordered form of cellulose Iβ.
Methods for measuring the crystalline
forms and crystallinity of cellulose

• X-ray diffraction
• Solid state Nuclear Magnetic Resonance (NMR) spectroscopy
specifically: cross-polarization magic angle spinning (CP-MAS) 13C NMR

Most applied methods, generally regarded as the most reliable

• Electron diffraction
• Neutron scattering
• IR spectroscopy
• Raman spectroscopy
Native cellulose:
Cellulose Iα
Cellulose Iβ
Cellulose I crystal
Cellulose chains form sheets which are connected with each other
Radial cross section of a cellulose Iβ crystallite:

6×6 model (not confirmed!)


→ 36 cellulose chains

Within the sheets:


hydrogen bonds

Between the sheets:


van der Waals bonds

Cross sectional image taken from: Gross and Chu J. Phys. Chem. B 2010, 114, 13333.
Cellulose I: main hydrogen bonding
patterns
Main hydrogen bonds:
Sheet in cellulose I
• 3→5 intramolecular bond
1 lends rigidity to the cellulose
2 chain
5

• 2→6 intramolecular bond


3
6 6 3
4
• 3→6 intermolecular bond
5 5 keeps the sheets together

NOTE: Cellulose chains run


6 3 parallel in cellulose I crystal
Distinction: cellulose Iα and cellulose Iβ
Two forms of native crystalline cellulose exist: Iα and Iβ.
Atalla and Vanderhart Science 1984, 223, 283.

Crystallographic details in 1Å
resolution (cellulose Iα ja Iβ):

Nishiyama et al.
J. Am. Chem. Soc. 2002, 124, 9074.
J. Am. Chem. Soc. 2003, 125, 14300.

Iα: one chain triclinic Iβ: two chain monoclinic


- dominant in, e.g., - dominant in higher plants
bacterial cellulose (e.g. wood, cotton)
and algae
Distinction: cellulose Iα and cellulose Iβ

Baker et al. J. Struct. Biol. 1997, 119, 129.


Distinction: cellulose Iα and cellulose Iβ

• Cellulose Iβ is the predominant form in higher plants (wood, cotton


etc.) and tunicate (cellulose in tunicate animals)

• Cellulose Iα is the predominant form in algae and in cellulose emitted


by microbes (bacterial cellulose)

NOTE: Cellulose Iα and cellulose Iβ ALWAYS coexists with each other in


nature, usually within the same microfibril.
Regenerated cellulose:
Cellulose II
Cellulose II
NATIVE
FORM Regeneration or
alkali treatment
Cellulose Iα
Cellulose II
Cellulose Iβ

“Man-made” form of cellulose


Preparation by: - dissolving the cellulose / regeneration
- swelling in concentrated alkali (e.g. > 10% NaOH)

Silk-like texture of cellulose II materials means that they


are widely applied in textile industry.
Distinction between cellulose I and II

Cellulose
(a) cellulose II Cellulose
(b) cellulose IIII

1
2 2
5 6

3
3 3
6 6 3
4

5 5 5
5 2
6

6 3

Hydrogen bonding patterns are different.


Distinction between cellulose I and II
Cellulose chain has a direction:
Reducing
OH OH end group
Anhydroglucose
O
OH O OH
O HO
HO HO O H
O O
HO HO n /2 OH
OH O OH
OH OH
Non-reducing end
Reducing end
Cellobiose

Cellulose I Cellulose II

Parallel Anti-parallel
Distinction between cellulose I and II
Cellulose II
Preparation by: - dissolving the cellulose / regeneration
- swelling in concentrated alkali

How is it possible for the cellulose chains to transform from


parallel to anti-parallel without dissolution?

Cellulose I Cellulose II

swelling

Parallel Anti-parallel
Distinction between cellulose I and II
• Cellulose crystals in parallel microfibrils run in opposite direction
• NaOH swells the crystals
• Anti-parallel arrangement is thermodynamically more favourable than
parallel arrangement
→ HYPOTHESIS:
Cellulose chains in parallel microfibrils intermingle and form new
antiparallel crystals

Cellulose I Cellulose II

swelling

Parallel Anti-parallel
Evidence for antiparallel cellulose II
Single cellulose I microfibril

CELLULOSE I

Two cellulose I microfibrils


Swelling in NaOH (aq)
(i.e. mercerization)

CELLULOSE II

Single cellulose II microfibril


Kim et al. Biomacromolecules 2006, 7, 274.
Evidence for antiparallel cellulose II
Dissolved end-functionalized cellulose:

Exposure to gold surface/


regeneration Diffraction pattern
Sulphur binds with gold. of cellulose I

Au
→ Forced parallel orientation
Yokota et al. Adv. Mater. 2007, 19, 3368.
Cellulose III
Conversion into cellulose III

NATIVE
FORM Regeneration or
alkali treatment
Cellulose Iα
Cellulose II
Cellulose Iβ
IRREVERSIBLE

Liquid NH3 or REVERSIBLE


diamine treatment

Cellulose IIII Cellulose IIIII

Parallel chains Anti-parallel chains

Hess and Trogus Ber. 1935, B68, 1986–1988.


Conversion: cellulose Iβ → cellulose III
Cellulose Iβ Cellulose IIII

Ammonia-cellulose I complex

NH3 (l)
NH3 removal
(e.g. evaporation)

Wada et al. Macromolecules 2006, 39, 2947.


Distinction: cellulose I and cellulose III
Cellulose Iβ Cellulose IIII

• Hydrogen bonds only between • Hydrogen bonding also between


cellulose molecules within the the sheets
sheets
• van der Waals bonds between the
sheets

Wada et al. Macromolecules 2006, 39, 2947.


Distinction: cellulose I and cellulose III
Cellulose Iβ Cellulose IIII

Chundawat et al. J. Am. Chem. Soc. 2011, 133, 11163.


Reversibility of cellulose III
• Cellulose III can be converted back to its starting material

Cellulose Iα
Cellulose II
Cellulose Iβ
Hydrothermal treatment
at ~160°C
or
Thermal treatment at >200°C

Cellulose IIII Cellulose IIIII

Parallel chains Anti-parallel chains

Chanzy et al. Carbohydr. Res. 1987, 160, 1.


Wada et al. Macromolecules 2001, 34, 3271.
Conversion: cellulose III → cellulose Iβ
Black: cellulose IIII
Grey: cellulose Iβ

Wada et al. Macromolecules 2001, 34, 3271.


Cellulose IV
Conversion: cellulose III → cellulose IV
NATIVE
FORM Regeneration or
alkali treatment
Cellulose Iα
Cellulose II
Cellulose Iβ

Liquid NH3 or
diamine treatment

Cellulose IIII Cellulose IIIII

Heat treatment in
glycerol at 260°C

Cellulose IVI Cellulose IVII

Hutino and Sakurada Naturwissenschaften 1940, 28, 577.


Conversion: cellulose III → cellulose IV
– recent evidence
Recent evidence from FT-IR spectroscopy, solid state NMR spectroscopy,
X-ray diffraction and diffraction simulations:

Wada et al. Biomacromolecules 2004, 5, 1385.


Newman Cellulose 2008, 15, 769.

• Credible proof that cellulose IVI is not a genuine allomorph


• Cellulose IVI is seen as a distorted form of cellulose Iβ
Consequences of the different
crystalline forms
Stiffness properties

Form Elastic modulus


Cellulose I 138 GPa
Cellulose II 88 GPa
Cellulose IIII 87 GPa
Cellulose IIIII 58 GPa

The values are estimates for pure crystalline forms.

• Native cellulose I is decidedly stiffer and stronger than the “man-made” forms

Nishino et al. J. Polym. Sci. B 1995, 33, 1647.


Enzymatic hydrolysis: cellulose I vs. II

Hydrolysis of
cellulose II is much
faster and proceeds
to a greater extent
than that of
cellulose I.

NOTE: Cellulose II hydrate is a form of cellulose II with water molecules inside


its crystalline lattice.
Wada et al. Polym. Degrad. Stabil. 2010, 95, 543.
Enzymatic hydrolysis: cellulose I vs. II
vs. III
Glucose yield from enzymatic hydrolysis of different cellulose allomorphs

C-I: cellulose I C-I*: ammonia-cellulose complex


C-II: cellulose II C-III: cellulose III
AC: amorphous cellulose

Chundawat et al. J. Am. Chem. Soc. 2011, 133, 11163.


Morphology of cellulose:
Native cellulose microfibril
Microfibrils in the plant cell wall
Individual fibre Cellulose microfibril
10 μm Diameter: 2-20 nm
Cross sectional models of crystalline
cellulose microfibrils
6×6 (36) chain models

3×5 nm

3.8 nm
Ding and Himmel J. Agric. Food Chem. 2006, 54, 597. Sugiyama et al. Macromolecules 1991, 24, 4168.

• Based on Atomic Force Microscopy • Based on diffraction studies


• much debated model. • More accepted model
Cross sectional models of crystalline
cellulose microfibrils
24 chain model

• Based on diffraction, NMR, and FTIR spectroscopy data


• Debate is still open

Fernandes et al. PNAS 2011, 108, E1195


Cross sectional models of crystalline
cellulose microfibrils

Terminological note:

• 36 and 24 chain models refer to the smallest cellulose microfibrils,


such as those present in wood cells

• Often these smallest microfibrils are referred to as elementary fibrils

• In many species, the microfibrils are larger but they are multiplicates of
the elementary fibrils

• Often the microfibrils (or elementary fibrils) aggregate, forming larger


microfibrillar bundles
Crystallinity and width of microfibrils
Collected data from analyses of some native cellulose sources
Source Degree of Microfibril Microfibril
crystallinity width* width**
Algal cellulose >80 % 10 nm 10-35 nm
Bacterial cellulose 65-79% 5 nm 4-7 nm
Cotton linters 56-65% 5 nm 7-9 nm
Ramie 44-47% 5 nm 3-12 nm
Hemp 60% 3-5 nm 3-18 nm
Flax 56% 4-5 nm 3-18 nm
Dissolving pulp 43-56% 4-5 nm 10-30 nm

*) Deduced from X-ray diffraction (reflection from 110 lattice plane)


**) Deduced from transmission electron microscopy images

Zugenmaier, In: Crystalline cellulose and cellulose derivatives;


Springer: 2008; p. 208.
Crystalline-amorphous transitions in a
cellulose microfibril
Longitudinal order: fringed-fibrillar
model of native cellulose microfibrils
Crystallographic data presents evidence
that cellulose within microfibrils is not
totally crystalline.

Proposition:
cellulose runs through alternating
crystalline and “ amorphous” regions.
Fringed-fibrillar model of native cellulose
microfibrils
Not just one model

Hearle 1958 Dolmetsch 1968 Hess and Kiessig 1953

... to name but a few.


Fringed-fibrillar model – evidence?
Level-off degree of polymerization (LODP): cellulose depolymerizes rapidly
down to a certain level after which the degradation is minimal.

HCl HYDROLYSIS
5OC

18OC

40OC
80OC
LODP

Proposition: amorphous domains are hydrolyzed leaving the


crystalline domains intact.
Battista Ind. Eng. Chem. 1950, 42, 502.
Fringed-fibrillar model – evidence?
DP of ramie fibres after Small angle x-ray scattering
HCl hydrolysis pattern of untreated ramie

LODP

Crystallite length (i.e. length of crystalline domains)


by SAXS agrees with the level-off
degree of polymerization (LODP).

Nishiyama et al. Biomacromolecules 2003, 4, 1013.


Fringed-fibrillar model – evidence?
DP of ramie fibres after However,
HCl hydrolysis - the yield loss upon controlled
acid hydrolysis is very small
- SAXS pattern implies a very
short disordered region

Proposition: disordered regions


LODP
are merely 4-5 anhydroglucose
units in length.

Nishiyama et al. Biomacromolecules 2003, 4, 1013.


Fringed-fibrillar model – so what?
The disordered segments can be selectively targeted with acid hydrolysis.

H+
H+
H+ H+
H+

RESULT:
cellulose nanocrystals
LODP

LODP depends on the native


source of cellulose.

Rånby Discuss. Faraday Soc. 1951, 11, 158.


Fringed-fibrillar model – so what?
Alternating crystalline-amorphous regions explain well
the macroscopic mechanical properties of cellulosic materials.

The length and width of the crystalline domains


depend on the native source of the material.

Elastic properties of isolated cellulose


nanofibrils depend on their native source.

Page J. Pulp Paper Sci. 1983, 3, TR15.


Fringed-fibrillar model – implications?
• When you see data of the ”degree of crystallinity” of cellulose, its physical
meaning is unclear
• If the degree of crystallinity is, e.g., 64%, it does not mean that 64% of the
cellulose is crystalline and 36% is ”amorphous”
• Probably much of the material responsible for the ”amorphous” response
resides on the microfibril surface
• Cellulose I and cellulose II degrees of crystallinity should never be
compared with each other
• Systematic sets of data can be compared with each other if the crystalline
forms, the analytical method, and the raw materials are similar
Aggregation or coalescence of
cellulose microfibrils inside the cell wall
Appearance of a cellulose microfibril
Aggregates: 12-20 nm Individual microfibrils: ~3.5 nm
200 nm

TEM image of longitudinal


AFM image of a surface of
cross-section of chlorite
bleached birch kraft pulp;
delignified pine cell wall;
sample untreated.
freeze-dried and stained.
Heyn J. Ultrastructure Res. 1969, 26, 52.
Appearance of a cellulose microfibril
Aggregates: 12-20 nm Individual microfibrils: ~3.5 nm

TEM image of radial TEM image of longitudinal


cross-section of wood cell wall. cross-section of chlorite
Zimmermann et al. delignified pine cell wall;
J. Struct. Biol. 2006, 156, 363. freeze-dried and stained.
Heyn J. Ultrastructure Res. 1969, 26, 52.
Implications of microfibril aggregation

Felling/ Why is this important?


drying

Reduced surface area


Aggregation

Reduced accessibility

Schematic cross sections of microfibrils

Fewer reaction sites Difficulties to extract


cellulose nanofibrils

Suchy et al.
Biomacromolecules 2010, 11, 515.
Summary
• Cellulose exists in several crystalline allomorphs:
- cellulose Iα and Iβ (native forms)
- cellulose II (prepared regeneration or alkaline treatment)
- cellulose IIII and IIIII (prepared by liquid ammonia treatment)

• Cellulose allomorphs are physically different and they differ in reactivity

• Structure of cellulose microfibril is an unsettled issue (36 and 24 chain


models)

• Cellulose in microfibrils is not totally crystalline: the order is frequently


interrupted by longitudinal disorder (fringed-fibrillar model)

• Microfibrils have an inherent tendency to coalesce with each other

• Disorder and coalescence affect the macroscopic physical properties of


cellulosic materials