Bio-Nanohybrid Materials

Eduardo Ruiz-Hitzky, Margarita Darder, Pilar Aranda

Instituto de Ciencia de Materiales de Madrid, CSIC

Spain

2nd NANOSPAIN WORKSHOP, BARCELONA 2005

AIM OF THIS COMMUNICATION

-Overview of procedures to obtain biopolymer nanocomposites providedwith specific functionality

-Applications of such materials towards advanced devices

-Research results from our own laboratory

ORGANIC-INORGANIC HYBRID MATERIALS

• Grafting of organic groups

• Intercalation of organic compounds

• Sol-gel methods

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grafting of organosilanes on silicic surfaces

polymer & biopolymer

nanocomposites

EXAMPLES

self-templating synthesis of organosilicic compounds

Nanostructured Organic-Inorganic Functional materials have to be prepared by a combination of physical and chemical methods following the modus operandi characteristic of the procedures of molecular engineering.

Idealized representation of a “nanochemist” applying methods for deliberate modifications of the functional properties of a 2D solid material. The picture includes some laborers of theoretical formation studying physical processes into the solid.

E. RUIZ-HITZKY “Organic-Inorganic Materials: From Intercalations to Devices”. Chapter 2. In: P. Gómez-Romero, C. Sánchez, eds. Functional Hybrid Materials; Wiley-VCH VerlagGmbH, 2004.

STRUCTURAL & FUNCTIONAL NANOCOMPOSITES

COMPOSITE MATERIALSAre solids resulting from the combination

of two or more simple materials that develop a continuous phase

and a dispersed phasewhich together have a set of properties

that is essentially different from the components taken separately.

polymer metal

ceramic...

glass fibersilica...

NANOCOMPOSITE MATERIALS

Are composites in which the dispersed phase presents at least one dimension at the nanometric scale.

1-nano-DLayered solidsSilica, C…

3-nano-DFibres, tubes…2-nano-D

HYBRID NANOCOMPOSITES BASED ON ORGANIC POLYMERS AND LAYERED SOLIDS

Layer thickness1 nm

Layer length: 1000 nm

Intercalated Delaminated(exfoliated)

Polymer

Micro-composite

EXAMPLES OF POLYMER-LAYERED SOLID NANOCOMPOSITESPossibilities:• inorganic host: non-charged, positively or negatively charged layers,

insulator, semiconductor, redox-sites, …• polymers: insulators, conducting, ionomers, polyelectrolytes…

Examples: Host

layered silicates (clays) (montmorillonite,. ..)

layered double hydroxides (MgAl, ZnAl, ZnCr,…)

phosphates & phosphonates (α-ZrP,…)

metal oxides (V2O5·nH2O, CoO2, MoO3,..)

metal chlorides & oxychlorides (FeOCl, α-RuCl3,…)

metal chalcogenides (MoS2, NbSe2,…)

metal phosphochalcogenides (MnPS3, CdPS3,…)

graphite oxide

polymer

PVA, PS, PSS, PMMA, PAN, PEG, PEO, PANi, PPy

PSS, PVS, PAA, PANi

PPy, PANi,…

PVP, PPV, PANi, PPy, PTh, PEG, PEO

PVP, PPy, PANi, PTh, PEO

PEO, PANi

PEO, PANi,

PEO, PANi

LAYERED SILICATES (CLAYS)Natural or synthetic phyllosilicates. Type 2:1 charged silicates (montmorillonite, hectorite,..)

electrical neutrality is kept by exchangeable interlayer cations

TWO tetrahedral layers of MIV (eg. Si) which are co-ordinated by oxygen anions to MIII (eg. Al) in ONE octahedral layer

a large variety of characteristics due to:- nature of cations MII ,MIII and MIV

- nature of the interlayer cation

1.20 nm1.20 nmsubstitution of MIII cations by MII

cations in the octahedral layer and/or substitution of MIV cations by MIII cations in the tetrahedral layer

overall negative charge

LAYERED DOUBLE HYDROXIDES (LDHs)LDHs are synthetic or natural solids with a layered structure similar to that exhibited by natural Mg(OH)2, brucite.

substitution of MIII cations into the brucite-like hydroxide layer

overall positive charge

electrical neutrality is kept by exchangeable interlayer anions

layers of MII and MIII cations which are octahedrally co-ordinated by six oxygen anions, as hydroxides

a large variety of characteristics due to:- nature of cations MII and MIII

- nature of the interlayer anion

Characteristics:• high charge density in the layers• anion-exchange properties: ion-exchange capacity 2-3 times greater than clays

Bio-nanocomposites : State of the Art & Examples

Bio-nanohybrids from collagen/alginate and hydroxyapatite: biomaterials for bone tissue engineering and drug delivery:

(Sotome et al. Mater. Sci. Engineering C, 2004, 24, 341-347)

newly formed bone retained the original size of the bio-nanocomposite implant

Negatively charged DNA has been intercalated in a LDH matrixLDH works as a nonviral vector to transfer the DNA into the cells:

(Choy, et al. Angew. Chem. Int. Ed., 2000, 39, 4042-4045)

Uptake by thecell

(endocytosis)

DNA-LDHhybrid

Slow LDH removalin the lysosome

Slow release of DNA

OBJECTIVES

The development of bio-nanocomposites based on the intercalation of natural polysaccharides into

layered solids (clays & LDHs)

Reinforcement of biopolymersTuneable ion-exchange behaviour

•Application of the bio-nanocomposites as activephase of potentiometric sensors

•Controlled release of electrically charged drugs

CHITOSAN-CLAY BIO-NANOCOMPOSITESChitosan: bio-polymer constituted by saccharide units (is a copolymer of glucosamine and N-acetylglucosamine).

OHO O O

O OHO O O

OO HO HO

OH

NH3+

NH3+

OH

OH

OH

NH3+

NH

CH3OCH3COO-

CH3COO- CH3COO-

In acid solutions (acetic acid, 1%) as a positively charged polymer (-NH3

+, pKa = 6.3), which is adsorbed on smectite clay suspensions.•Ion-exchange process

•Polymer-salt intercalation

M. DARDER, M. COLILLA, E. RUIZ-HITZKY Biopolymer-clay nanocomposites based on chitosan intercalated in montmorillonite Chem. Mater. 15, 3774-3780 (2003)

ANIONIC POLYSACCHARIDES LDH BIO-NANOCOMPOSITES

OO

OO

O

OO

O O

O-

O

OH

OHO-O

OH

O-

O

HO

HO

O-O

OH

OHOH

n

Alginate From brown algae

pKa guluronicacid units = 3.65

pKa mannuronicacid units = 3.38

O

O

OH

OH

OO

O

OSO3-

O

-O 3 S O

n

OOH

HO

O

O

CH3O

OHO

HO

O

OO

-O

n

pKa = 2.9

PectinFrom citrus fruits

iota-CarrageenanFrom red algae

anionic polysaccharides-LDH bio-nanocomposites

+ nNa+X-OHO O O

O OO O

OO HO HO

OH

NH3+ NH3

+OH

OH

HO

NH

CH3O

OH

NH3+

1.45 nm

Chit-NH3+ X-

2.09 nm

OHO O O

O OHO O O

OO HO HO

OH

NH3+

NH3+

OH

OH

OH

NH3+

NH

CH3O X-X-

Chit-NH3+ X- +

Hydrated Na+

cations

1.20 nm

CH3COO-

ions

OHO O O

O OO O

OO HO HO

OH

NH3+ NH3

+OH

OH

HO

NH

CH3O

OH

NH3+

1.63 nm

Pectin or alginate

Cl- anions

0.48 nm

+ + + +

+ + + +

+ nNa+Cl-1.32 nm

+ + + +

+ + + +

OOH

HO

O

O

C H3O

OHO

HO

O

OO

-O

n

+ + + +

+ + + +

+ nNa+Cl-

ι-Carrageenan

A

M+M+

-O3SOOH

OO

O

OSO3-

O OO

OH

-O3SOOH

OO

O

OSO3-

O OO

OH

+ nNa+X-OHO O O

O OO O

OO HO HO

OH

NH3+ NH3

+OH

OH

HO

NH

CH3O

OH

NH3+

1.45 nm

Chit-NH3+ X-

2.09 nm

OHO O O

O OHO O O

OO HO HO

OH

NH3+

NH3+

OH

OH

OH

NH3+

NH

CH3O X-X-

Chit-NH3+ X- +

Hydrated Na+

cations

1.20 nm

CH3COO-

ions

OHO O O

O OO O

OO HO HO

OH

NH3+ NH3

+OH

OH

HO

NH

CH3O

OH

NH3+

1.63 nm

Pectin or alginate

Cl- anions

0.48 nm

+ + + +

+ + + +

+ nNa+Cl-1.32 nm

+ + + +

+ + + +

OOH

HO

O

O

C H3O

OHO

HO

O

OO

-O

n

+ + + +

+ + + +

+ nNa+Cl-

ι-Carrageenan

Achitosan-montmorillonite bio-nanocomposites

M+M+

-O3SOOH

OO

O

OSO3-

O OO

OH

-O3SOOH

OO

O

OSO3-

O OO

OH

M. DARDER, M. COLILLA, E. RUIZ-HITZKY Biopolymer-clay nanocomposites based on chitosan intercalated in montmorillonite Chem. Mater. 15, 3774-3780 (2003)

M. DARDER, M. LOPEZ-BLANCO, P. ARANDA, F. LEROUX, E. RUIZ-HITZKY Bio-nanocomposites based on layered double hydroxides Chem. Mater. 17, 1969-1977 (2005)

Intercalation of chitosan in Na+-montmorillonite

+ nNa+ X-O

HO O OO O

O OOO HO HO

OH

NH3+ NH3

+OH

OH

HO

NH

CH3O

OH

NH3+

1.45 nm

Chit-NH3+ X- +

hydrated Na+ cations

1.20 nm

CH3COO- ions

Chit-NH3+ X-

OHO O O

O OO O

OO HO HO

OH

NH3+ NH3

+OH

OH

HO

NH

CH3O

OH

NH3+

2.09 nm

OHO O O

O OHO O O

OO HO HO

OH

NH3+

NH3+

OH

OH

OH

NH3+

NH

CH3O X-X-

Bilayer

Adsorption isotherm (323K)

Monolayer

0 4 8 12 16 200

50

100

150

200

equilibrium concentration/mmol L-1

adso

rbed

qua

ntiti

es/1

0-3 Eq

per

100

g

Theoretical anionic exchange capacity of 57 mEq/100g

ONE-POT SYNTHESIS OF BIO-POLYMER/LDH NANOCOMPOSITES

Co-organized assembly ortemplating co-precipitation:

LDH precursors+ polymer

Bio-nanocomposite

control of pH

TEMPLATE SYNTHESIS OF BIOPOLYMER-LDH NANOCOMPOSITES

Anionic biopolymersolution, under magnetic

stirring and N2 atmosphere

ZnCl2 + AlCl3solution

Dropwiseaddition of

Zn/Al solutionPeristaltic

pump

1.0 M NaOHsolution

pH-meter9.0 - 9.5

Controlledaddition of NaOH

automatic dispenser

“coprecipitation” or “co-organised assembly”

M. Darder, M. Lopez-Blanco, P. Aranda, F. Leroux, E. Ruiz-Hitzky Chem. Mater. 17, 1969-1977 (2005)

CHARACTERIZATION OF THE

BIOPOLYMER- NANOCOMPOSITES

X-ray diffraction

FTIR spectroscopy

Solid state 13C NMR spectroscopy

Scanning electron microscopy (SEM)

Thermogravimetry (TG) anddifferential thermal analysis (DTA)

Direct potentiometry

X-Ray Diffraction

0 5 10 15

Amount of chitosanin nanocomposites(mEq/100g clay): 24.7 44.1 76.6 145.3 173.8 180.8

Chitosan film

SWy-1

2.09 nm

2.04 nm

1.34 nm

2.00 nm

1.69 nm

1.45 nm

1.39 nm

1.20 nm

Inte

nsity

/ a.

u.

2θ

Monolayer

Bilayer

[Zn2Al(OH)6]Cl

0 10 20 30 40 50 60 70

0.15

1 (1

13)

0.15

4 (1

10)

0.20(018)

0.23(015)0.26

(012)

0.38(006)

0.76(003)

0.150.26

0.420.490.64

1.32

0.15

0.260.43

0.651.37

0.420.570.82

1.63

Inte

nsity

/ a.

u.

2θ / degrees

ι-carrageenan-LDH

pectin-LDH

alginate-LDH

Chitosan/montmorillonite

G4: 74.4 ppm 75.0 ppmA2: 69.2 ppm 69.6 ppm

Solid-state 13C NMR spectroscopy

O

O

OH

OH

OO

O

OSO3-

O

n

- O 3 S OA1

A2 A3

A4

A5

A6

G1

G2

G3

G5G4 G6

200 150 100 50 0

98.8

102.481.0

32.262.3

69.6

75.0

90.7

104.0

62.4

69.2

74.477.3

90.9

104.9

ppm

ι-carrageenan

ι-carrageenan-LDH

200 150 100 50 0 -50

23.4(CH

3)

56.4(C2)

60.6(C6)

22.9

56.459.5

74.7(C3)

82.7(C4-C5)

102.6(C1)

74.6

81.8

103.4

174.9 (C=O)

x4

x4

ppm

-NH3+NO3

-

-NH3+CH3COO-

• Anion-exchange behavior

13C NMR Spectroscopy

OHO

OH

NH

CH3O

O

NH3+

OH

HOO

OO C1 C2C3

C4 C1’C5

C3’

C4’ C5’C6’

C6

C7

C8

C2’

OHO

OH

NH

CH3O

O

NH3+

OH

HOO

OO C1 C2C3

C4 C1’C5

C3’

C4’ C5’C6’

C6

C7

C8

C2’

TREATMENT WITH NaNO3

Chitosan-clay bio-nanocomposite

Development of potentiometric sensors

PVC-membrane based electrodes

Bio-nanocomposite-modifiedPVC membrane

Silver wire

Internalreferenceelectrode(Ag/AgCl)

KCl 3M + AgCl sat

Internal referencesolution

Bio-nanocomposite-modifiedcarbon paste

Plastictube

Copper wire

Carbon paste electrodes(CPEs)

Chitosan-clay sensors development

The good mechanical properties of bio-nanocomposite avoid the use of binders as it is usual in CPEs or Epoxy Based Sensors

Chitosan-claynanocomposite

aqueous suspension

Addition of graphite powder

Disc of graphite-nanocomposite

Dried at50 ºC

Chitosan-clay-graphite

nanocomposite

Plastictube

Copper wire

Electrochemicalsensor

Chitosan-claynanocomposite

aqueous suspension

Chitosan-claynanocomposite

aqueous suspension

Addition of graphite powder

Addition of graphite powder

Disc of graphite-nanocomposite

Dried at50 ºC

Disc of graphite-nanocomposite

Dried at50 ºC

Chitosan-clay-graphite

nanocomposite

Plastictube

Copper wire

Electrochemicalsensor

Chitosan-clay-graphite

nanocomposite

Plastictube

Copper wire

Chitosan-clay-graphite

nanocomposite

Plastictube

Copper wire

Electrochemicalsensor

• Potentiometric response

-6 -5 -4 -3 -2 -1 00

40

80

120

160

200

E / m

V

log a / M

Potentiometric measurement of NaCl with sensors prepared from different nanocomposites

Nanocomposite with a chitosan monolayer

Positive slope

Potentiometric response to cations

Nanocomposite with a chitosan bilayer

Negative slope

Potentiometric response to anions

Chitosan-montmorillonite potentiometric sensorThe potentiometric response of a sensor based on the chitosan-clay nanocomposite with a chitosan bilayer is evaluated towards different anions

fig

-6 -5 -4 -3 -2 -1 0

80

120

160

200

240

280

320

360 Cl-

NO3-

SO42-

Cr2O72-

Fe(CN)63-

CH3COO-

F-

E / m

V

log aX- / M

M. DARDER, M. COLILLA, E. RUIZ-HITZKY Chitosan-clay nanocomposites: application as electrochemical sensorsAppl. Clay Sci. 28, 199-208 (2005)

0.0

-1.0

-1.5

NO3-

CH3COO- Cl- F-

SO4

2- Cr2O7

2-

Fe(CN)6

3-

-0.5

POTXNO

K −− ,3log

Selectivity coefficients

The sensor is 25 times more selective for nitrate than divalent anions

The sensor is 60 times more selective for nitrate than trivalent anions

Partial selectivity to monovalent

anions

Cross-sensitivity

NO3− ≈ CH3COO− ≈ Cl− >>> SO4

2− ≈ Cr2O72− >> Fe(CN)6

3−

LDH Bio-nanocomposite CPEs

E = 35.9 − 57.3 log aCl-

r = 0,9986

E = 213.5 + 22.2 log aCa2+

r = 0,998

-6 -5 -4 -3 -2 -1

90

120

150

180

210

240

270

300

E /

mV

log aX / M

ι-carrageenan-LDH

Zn2Al/Cl LDH

The AEC of the LDH has been reversed to

a CEC in the bionanocomposite

Sensor arrays controlled by Artificial InteligenceCombination of sensors with AI techniques give sophisticated devices, as artificial noses and tongues

Electrodes array

Reference electrode

Multi-channel box

Data adquisition and treatment

Data processing

Liquid sample

Application of Case-Based Reasoning (CBR) for multicomponent analysis

Applications

-water quality: drink water & industrial pollution

-Ions in biological fluids

-Automation in fertiirrigationM. COLILLA, C.J. FERNÁNDEZ, E. RUIZ-HITZKYThe Analyst, 127, 1580-1582 (2002)

Home-made “electronic tongue” entirely developed in our lab, using sensors based on bio-nanocomposites, applied to control the water quality

CONCLUSIONSCationic or Anionic polysaccharides form bio-nanocomposites by combination at the nanometric scale with layered silicates (INTERCALATION) or double hydroxides (CO-ORGANISED ASSEMBLY) resulting infunctional hybrid nanostructured materials with:

• Good mechanical properties

• Controlled ion-exchange behaviour

These results open a way to prepare novel bio-nanohybrids provided of structural or functional properties.

FUTURE WORK

The use of other layered host solids (phosphates, phosphonates, chalcogenides, etc.) & bio-polymers (polypeptides, nucleic acids, etc..)

Projects Financed by

CICYTMAT2003-06003-C02-01

Comunidad Autónoma de Madrid07N / 0070 / 2002

Junta de AndalucíaIFAPA – 2002.000890