Department of Applied Physics,

The Hebrew University of Jerusalem,

Israel

Yuri Feldman

1

Water in heterogeneous/biological systems

Interfacial water—from the ordered structures to the

single hydrated shell

2

Dielectric spectroscopy is sensitive to relaxation processes

Types of polarization

Electric Field

Orientation polarization:

Ionic Polarization:

+

-

--

- +

++ --

+

+-

-

+

++-

--

--

-

-

-

++ + +

+

+

+

+ +

Electric Field

+

Deformation polarization

Introduction-Dielectric Spectroscopy

~

32

Dielectric spectroscopy is sensitive to relaxation processes

Types of polarization

Electric Field

Orientation polarization:

Ionic Polarization:

+

-

--

- +

++ --

+

+-

-

+

++-

--

--

-

-

-

++ + +

+

+

+

+ +

Electric Field

Deformation polarization

~

Introduction-Dielectric Spectroscopy

4

Dielectric spectroscopy is sensitive to relaxation processes

in an extremely wide range of characteristic times ( 10 5 - 10 -12 s)

Broadband Dielectric Spectroscopy

Porous materialsand colloids

Clusters Single droplets and pores

Glass forming liquids

Macromolecules

10-210-4 0 102 104 106 108 1010 1012

Time Domain Dielectric Spectroscopy; Time Domain Reflectometry

f (Hz)

10-6

Water

ice

Dielectric response on mesoscale

Dielectric Response in Biological Systems

5

Broadband Dielectric Spectroscopy

Cells

Time Domain Dielectric Spectroscopy

f (Hz)H

H3N+ — C — COO-

R

Ala Asp Arg Asn

Cys Glu Gln His

Ile Leu Lys Met

Phe Ser Thr Trp

Tyr Val

1031020 105 106 107 108 109 101010410110-1 1011

P-

N+Head group

region

Lipids

Proteins

Water

- Dispersion

DNA, RNA

-Dispersion - Dispersion - Dispersion

Tissues

ice

Amino acids

+ =

Water as a marker in the dielectric spectroscopy measurements

1.8-3 D

(large dipole moment)

1031020 105 106 107 108 109 101010410110-1 1011

Decreasing temperature/

Confined conditions

Water is the “contrast” in dielectric measurements!!!

7

Water as a marker in the dielectric spectroscopy measurements

8

10 -110 1

10 3

10 510 7

Frequency [Hz]

10-4

10-2

100

102

Perm

ittivity''

[]

-200 0 200 400

Temperature [°C]

10 1

10 3

10 5

10 7

Frequency [Hz]

10-4

10-2

100

102

Pe

rmitt

ivity

'' []

0100

200300

Temperature [°C]

Water as a marker in the dielectric spectroscopy measurements

Hydrated sampleDehydrated sample

1031020 105 106 107 108 109 101010410110-1 1011

Decreasing temperature/

Confined conditions

Water is the “contrast” in dielectric measurements!!!

9

1) Silica glasses

a. 62.6% SiO2, 30.4% B2O3,7%Na2O

b. 70% SiO2, 23% B2O3, 7% Na2O,

2) The study of confined water dynamics in clay

minerals with different doped ions (K, Co, Ni)

a. Montmorillonite

b. Kaolinite

Clays and Clay Minerals (2014), Vol. 62, pp. 62–73

Water in non organic systems:

Microporous and Mesoporous Materials

58 (2003) 237–254

PRL (2010) Vol. 105, pp. 037601-4

10

L-defects D-defects OH- OH3+

Orientation defectsIonic defects

Dielectric relaxation in ice and water

Si

OO

O

10 -110 1

10 310 5

10 7

Frequency [Hz]

10-4

10-2

100

102

Perm

ittivity''

[]

-200 0 200 400

Temperature [°C]

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Local motion

Inter center

motion

Porous glasses: Relaxation Mechanism

12

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

Si

OOO

10 -110 1

10 310 5

10 7

Frequency [Hz]

10-4

10-2

100

102

Perm

ittivity''

[]

-200 0 200 400

Temperature [°C]

Si

OOO

Si

OOO

Si

OOO

Porous glasses: Relaxation Mechanism

𝑴 =

𝑖

𝑁

𝑒𝑖𝒓𝑖

10 1

10 3

10 5

10 7

Frequency [Hz]

10-4

10-2

100

102

Pe

rmitt

ivity

'' []

0100

200300

Temperature [°C]

10 -110 1

10 3

10 510 7

Frequency [Hz]

10-4

10-2

100

102

Perm

ittivity''

[]

-200 0 200 400

Temperature [°C]

SamplesHumidity h, %

II 0 .63

A 1 .2

B 1 .4

D 1 .6

C 3 .2

III 3 .39

I 3 .6

Porous glasses

14

A - 50 kJ/mol

B - 42 kJ/mol

C - 67 kJ/mol

D - 19 kJ/mol

Ice - 60 kJ/mol

15

Porous glasses

4.6 4.8 5.0 5.2 5.4 5.6 5.8

10-7

10-6

10-5

10-4

10-3

3.5 3.7 3.9 4.1 4.3 4.5 4.7 Sample

A

B

C

D

Ice

, [

s]

1000/T, [K-1]

170 180 190 200 210 2200.01

0.1

1

A

B

C

D

Temperature, [K]

-17 -16 -15 -14 -13 -12 -11 -10 -9 -8

0.36

0.39

0.42

0.45

0.48

0.51

0.54

0.57

0.60

0.63

A

B

C

D

I

II

III

ln()

A - 50 kJ/mol

B - 42 kJ/mol

C - 67 kJ/mol

D - 19 kJ/mol

Ice - 60 kJ/mol

16

2.42.62.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.8

10-8

10-7

10-6

10-5

10-4

10-3

10-2

A

B

C

, [s]

1000/T, [K-1]

II process

I process

Porous glasses

17

Aluminosilicates: Montmorillonite Ni

10-1101

103105

107Frequency [Hz]

10

-310

-110

110

310

5

Pe

rmittivity''

-200 -100 0 100 200 300

Temperature [°C]

3

2

1

5.6 5.8 6.0 6.2 6.4 6.6

10-6

10-5

10-4

10-3

10-2

[s]

1000/T

Co Co2

Ni Ni2

Cu Cu2

18

2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5

10-9

10-8

10-7

10-6

10-5

10-4

10-3

10-2

10-1

II.2

II.1

I.2

I.1

Montmorillonite,

s

1000/T

I K

I K2

I Co

I Co2

I Ni

I Ni2

I Cu

I Cu2

II K

II Co

II Co2

II Ni

II Ni2

II Cu

II Cu2

Aluminosilicates: Montmorillonite Ni

19

From non organic to organic systems

Using the water as a marker in porous system we can find:

1) The dynamics of water in the regime of the tight confinement

2) The influence of the various ions on the water dynamic

3) Structural properties of the sample: percolation cluster, fractal dimension and

porosity

In organic systems additional effects appear:

Specific structure of the

protein surface

Concentration of the

hydrophilic centers in

one place

Anisotropic properties.

Ordered structures

Inert interfacial water as apposed to

the actively solvating molecule

150200

250300

350

10-1

101

103

105

0

2

4

6

MidT

Die

lectr

ic lo

sse

s, "

log(f)

Temperature, [K]

LowT

Percolation

Saddle-like

Dielectric Responses of the confined water are similar

in non organic and organic systems

10 -110 1

10 310 5

10 7

Frequency [Hz]

10-4

10-2

100

102

Perm

ittiv

ity''

[]

-200 0 200 400

Temperature [°C]

[Clays and Clay Minerals

(2014), Vol. 62, pp. 62–73]

PCCP (2016),

V.18, pp. 10992-

10999

Microporous and Mesoporous

Materials 58 (2003) 237–254

Hydrated

proteins

(Lysozyme)

Porous glasses

Clays

21

Protein functionality

Protein structure

Protein hydration

shell

22

Water in organic systems. Hydration shell dynamics in proteins:

1) Hydration shell dynamics of a fibril protein

(Collagen type 1).

2) Hydration shell dynamics of a globular protein

(Lysozyme).

3) Hydration shell dynamics of a ring like protein

(Phycocyanin).

23

Introduction to collagen: General structure

Collagen is a protein consisted of (X-Y-Gly)n repeating sequences

Gly is the Glycine X or Y is the Proline or 4-HydroxyProline

HO

Steric effect stabilizes the

extended nature of the

individual chains

Hydrogen bonds (Gly: NH∙∙∙O=C: X)

leads to triple helixes organization Stable triple

helixes structure

24

Introduction to collagen: Water bridges

Water molecules embed into triple helixes

creating water bridges, that stabilizes collagen

structure [J. Bella et al Science, 1994]

Water bridges appear as a

surface bridges surround the triple helixes.

25

The problem: What is the differences

between hydration shell of globular

and fibrillar proteins

Introduction to collagen: Water bridges

The hydration shells connect between each

other creating stabilized collagen filaments

[K. Kawahara et al, Biochemistry, 2005]

26

Dielectric measurements of the hydrated collagen powders

S. A. Lusceac, et al Proteins and

Proteomics, 2010

3D plot of dielectric losses of the hydrated

collagen powders

h=0.16 =0.26 =0.33

Low temperature process

27

Main processIce process

S. A. Lusceac, et al J. Non-Cryst.

Solids (2011)

Dielectric measurements of the hydrated collagen powders

Ice process appears at high hydration level h>0.4

1. Main process doesn’t depend on the shape of the protein and is observed in

the various proteins.

2. It is attributed to the large-angle jumps of the water molecule

3. The present models are ignored the excess wing at the high frequency

S. A. Lusceac, et al Proteins and

Proteomics, 2010

28

New approach in the data description: Excess wing explanation

The proposed large-angle jumps can be corresponded with the migration of the H-bond

network defect [I. Popov, A. Puzenko, A. Khamzin and Y. Feldman PCCP, 2015]

*

12 *2

0

( )( )

16

or

B

i nq r

k T

2 2( ) /or

or hr t t

1*2 ( ) ~

or

orr i

*( )

1or

ori

If we take into account an ionic defect

Orientation defect

Ionic defect

*

12

2 *2

10

( )

1 ( )6

i i i

iB

in q r

k T

*

1( )

1or ion

or ioni i

29

*

1( )

1or ion

or ioni i

Collagen data treatment

30

Relaxation in ice

log τ

1000/T

Orientation

defect

Ionic

defect

I. Popov, A. Puzenko, A. Khamzin

and Y. Feldman PCCP, 2015

???

Orientation

Ionic

Additional delay due to

correlation with L-D defects

31

Relaxation in ice

log τ

1000/T

Independent defect migration

H3O+ defect

L- defect Blockage of the ionic defect

H3O+ defect

L- defectL-D defects

ionic defects

The low-temperature dynamic crossover in dielectric

relaxation behavior of ice Ih

Popov I., Lunev I., Khamzin A., Greenbaum A, Feldman Yu.

(2017, Paper in preparation)

32

Relaxation in ice

K.Goto et al Japanese J.

of Applied Phys. (1986)

pp.351-357

At low temperature the crack

appearance leads to the suppression of

the ionic defect and transition back to

the mechanism of the orientation

defect.

33

Collagen data treatment

H3O+

H3O+

Relaxation by ionic defect cannot be faster than relaxation by orientation defect

Hydration water ~ Distorted ice

Dehydrothermal Treatment:

34

After Heating and Keeping the

collagen at 120 0C for 30 min, proton

hopping mechanism slows down

Chains tilting prevents the

formation of the long range

water structure.

Relaxation occurs in local

compartments, where

contribution of the L-D and

ionic defect are comparable

Lysozyme: Surface charge plot[Christopher D. Cooper at el. 2013]

Charged active center

Lysozyme

35

~20Å

BDS Measurements: Lysozyme h=0.28 and h=0.16

150200

250300

350

10-1

101

103

105

0

2

4

6

MidT

Die

lectr

ic lo

sse

s, "

log(f)

Temperature, [K]

LowT

Percolation

Saddle-like

(a)

The typical dielectric spectra

[Khodadadi and A.P.Sokolov 2015] 36

37

5.0 5.5 6.0 6.5 7.0 7.5

-7

-6

-5

-4

-3

-2

-1

0 h=0.16

h=0.28

1000/T, [K-1]

log[

], [s]

T=184K

Ea=30kJ/mol

Ea=50kJ/mol

The typical dielectric spectra

[Khodadadi and A.P.Sokolov 2015]

4.5 5.0 5.5 6.0 6.5 7.010

-8

10-7

10-6

10-5

10-4

10-3

10-2

10-1

Lysozyme (h~0.37)

Myoglobin (h~0.33)

log[

], [s]

1000/T, [K-1]

Swenson and CerVeny 2015

BDS Measurements: Lysozyme h=0.28 and h=0.16

Temperature

solid like

behavior

liquid like

behavior

non-

correlated

system

Entropy and

Static Dielectric Permittivity

E=0

Dielectric Polarization and Entropy of the System

2 2

0 0

( ) ( )( ) (T) (T)

8 8

s T E T ES T S S

T T

ሻΔε(𝑇

Solid (Ordered system) Liquid (Disordered system)

E=0 E≠0

Decrease ordering Increase ordering

( )0s T

T

( )0s T

T

Τ𝑑Δε 𝑑𝑇>0 tendency tosolid-like dipole orientation

Τ𝑑Δε 𝑑𝑇<0 tendency points to a liquid-like behavior.

Tm of the extremum of 𝛥𝜀(T), the reorientation transition temperature.

T<155K 155K<T<187K T>187K

Short-range antiparallel orientation of the water dipoles, typical for the amorphous system

Tendency to solid like behavior

Tendency to liquid like behavior

39

BDS Measurements: Lysozyme h=0.28 and h=0.16

0 10 20 30 40

120

160

200

240

280

320

Tem

pera

ture

[K]

Time [minutes]

140 150 160 170 180 190 200 210 220

155 K

Cooling with no annealing

H

eat F

low

, [W

g-1]

Temperature, [K]

Exo 155K

DSC Measurements: Lysozyme h=0.2

150 175 200 225

Heat

Flo

w

Temperature, [K]

Exo

40

Protocol

140 150 160 170 180 190 200 210 220

155 K

Cooling with no annealing

Cooling with annealing

H

eat F

low

, [W

g-1]

Temperature, [K]

Exo

186.5K

155K

Lysozyme: Surface charge plot[Christopher D. Cooper at el. 2013]

Charged active center

Explanation of the Results

41

~20Å

∆𝑻𝒇(𝑹ሻ = 𝑪𝑮𝑻/(𝑹 − 𝒓ሻ

∆𝑇𝑓 = 𝑇𝑓 𝑏𝑢𝑙𝑘 − 𝑇𝑓 𝑝𝑜𝑟𝑒 - The depression of the confined water melting temperature.

𝐶𝐺𝑇-The Gibbs-Thompson Coefficient. R - The radius of the pore. r - The thickness of a liquid-like layer at the relevant melting temperature.

42

Gibbs-Thompson Fit for Silica Nano pores

0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.50

20

40

60

80

100

GT equation

MCM-41 (Findenegg, G. H. et al, 2008)

MCM-41 (Schmidt, R. et al, 1995)

SBA-15 (Findenegg, G. H. et al, 2008)

[

R, [nm]

𝒓𝑹

4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5

-10

-9

-8

-7

-6

-5

-4

-3

-2

-1

0 Hydrated silica nanopores

Hydrated lysozyme

log[

], [s]

1000/T, [K-1]

43

Relaxation Times of Hydrated Lysozyme and Confined Water in Silica Nanopores

20Å

Silica nanopore with pore diameter equaled to 20A

Lysozyme with diameter cavity inside equaled to 20A

Crossover Tc

[Eliodoro Chiavazzoet al 2014] [Christopher D. Cooper at el. 2013]

Glass-state water

T=155K

Temperature, [K]

T=184K

Glass solution

T=155K:

1) Glass transition of the

solution

2) Onset of the crystallization

Nucleation growth

of the ice particles

….Gradual

growth of the ice

particles…..

T=184K:

Phase Transition of the

ice inside the pore.

Ice

melting…

Ice particle inside

the pore

The Model

5.0 5.5 6.0 6.5 7.0 7.5

-7

-6

-5

-4

-3

-2

-1

0 h=0.16

h=0.28

1000/T, [K-1]

log[

], [s]

T=184K

Conclusions

45

2) Water can be considered as a marker or a contrast

in Dielectric Spectroscopy. The structural and

dynamical properties of the matriex can be studied via

its hydrating.

3) Defect migration model can be used as an universal

approach in description of the dynamic of the

hydration process.

1) The morphology, dynamics and the dielectric properties of the

matrixes will have an influence on the nature of the relaxation of

hydrated water.

I would like to thank my PhD student Mrs Yael Segev,for the

generation of the data and the deep discussion of obtained

results.

I would like also to appreciate Dr. Paul Ben Ishai, Dr Anna

Greenbaum, Dr. Ivan Popov and Dr. Evegenia Levy for their

extremely helpful contribution to this work

This work has been supported by the Israel Science

Foundation (ISF) (Grant No. 465/11);

46

Acknowledgements

47

Acknowledgements