Electron Paramagnetic Resonance (EPR) Electron Spin Resonance (ESR)

Electron Paramagnetic Resonance (EPR)
= Electron Spin Resonance (ESR)
I. Origins of EPR: Zeeman Interaction
II. Experimental considerations, Spectral presentation
III. Anisotropy and Spin-Orbit Coupling Components
IV. Hyperfine and Superhyperfine Splittings
V. Uses & Applications

A. Inorganic Spin Distribution
B. d-orbital splittings & EX Energies
C. Bioinorganic examples
D. Spin Labelling
© 2012, K.S. Suslick
Survey of Spectroscopic Techniques

cm-1 108 107 105 104 103 102 10 1 0.1 0.01 0.001
thz
mag moment
of nuclei
1
E R
Paramagnetic
Most substances do not contain paramagnetic species
and are hence EPR silent
Disadvantage Advantages
Fewer accessible systems 1) Easier to tell the source of EPR.
2) Introduction of “spin labels”
O
O
S S
N
N O
OH
O H2N
+ Protein-SH
S
S
Protein
N

O
EPR Pros & Cons
What EPR can tell us:

• What types of paramagnetic species are present
• What is the local structure/symmetry of these species
• What is the nature of the wavefunc0on containing the unpaired spin
(i.e., where are the unpaired spins localized ).
• For organic radicals, sub-µM; for TM complexes, mM to 10 µM;
300 µL volume. i.e., 10-1000 X more sensitive than NMR
Major disadvantages:
• Typically requires odd integer spins (i.e., S = 1/2, 3/2, 5/2, 7/2, etc…)
• Resolution is not as good as in NMR (broad features)

• Often requires very low temps for good resolution (10 K typical)
2
The Spin Hamiltonian
kinetic energy, e─ - nuclear attraction, e─ - e─

repulsion, nucl.-nucl. replusion, ligand field
“spin Hamiltonian”
Nuclear Zeeman (NMR)
Electronic Zeeman (EPR)
Zero Field Splitting (ZFS) from

spin-orbit coupling of angular mom.
Hyperfine Coupling of e─ & nuclear

moments (NMR & EPR)
Zeeman Interactions: Energy Splittings

→
1. An unpaired electron has a magnetic moment: µe
2. In an applied magnetic field, H0, along the z direction
classically,
3. For one free electron: cf. EI = - mIħH0 (nucleus)

define: -eħ =  ge
 B = Bohr magneton

= 9.27×10−24 J/T
3
4. For one free electron, the selection rule:
∆ ms = ± 1 electric dipole forbidden, mag dipole allowed only.
5. Energy difference: gβ(+½)H - gβ(-½)H = gβH = h
 B = Bohr magneton

= 9.27×10−24 J/T
Zeeman Interactions: Energy Splittings for 1 e-
4
Microwave frequency is fixed by the generator (“Klystron”),
so the magnetic field (H or B) is “scanned” until resonance found:

Microwave frequency is fixed by the generator (“Klystron”),
so the magnetic field (H or B) is “scanned” until resonance found:
5
The g-value
In the case of a free electron in

space, the e- is the only source
Ezee = - μe H0 = geemSH0
of magnetism in the sample:
ge 
In the case of an e- in a molecule,

SOC (spin-orbit coupling constant Ezee = - (μe + aλ) H0
) adds or subtracts orbital angular
momentum to that of free electron = gobsemSH0
and we lump that into “gobs” or “g”:
When unpaired electron couples to

1) Empty orbital (e.g., d1), g < ge
2) Occupied orbital (e.g., d9), g > ge
Generally 1.3 < g < 9
The g-value
Energy of the Zeeman transition (and hence the “g-value”) is determined
at a fixed microwave frequency with H0 being scanned. The “g-value”
is a unique property of the molecule as a whole and independent of any
electron – nuclear spin interactions.
h  (GHz ) 714.48  (GHz )

g  0.07145 
H H (T ) H ( gauss )
• For most organic radicals, g ≈ ge

• For transition metals, large deviations from ge possible
• g can be measure to high accuracy (±0.0001)
• g is the “chemical shift” equivalent of EPR
• g depends on structure of radical, excitation energies, strengths of
spin-orbit couplings
Note for later: g is anisotropic and not a scalar but a tensor.
6
What is the origin of g‐values diﬀerent from 2.0?
 Deviations from g = 2.0 due primarily to Spin‐Orbit Coupling. SOC constant, ζ,
is metal‐ion dependent: can be small (150 cm‐1; Ti3+) or large (830 cm‐1; Cu2+).
 An electron acquires orbital motion so to favorably experience the magnetic
field of a nucleus, hence lowering its overall energy.
 Specifically, an electron in an orbital (e.g., xy), can, via SOC, gain orbital angular
momentum via mixing with other orbital trajectories defined by rotational
operators. Hence, an electron in xy can gain xz character via rotation about X,
where such a rotation has a barrier, Δx.
 For an electron that rotates into an empty orbital, the "current" created by the
rotating electron leads to a magnetic field that opposes the applied magnetic field
B0. This requires a larger applied field to achieve resonance, and hence a lowering
of g.
 For an electron that rotates into an occupied orbital, the sense of the current
created by the rotating electron leads to a magnetic field that aligns with the
applied magnetic field B0. This requires a smaller applied field to achieve
resonance, and hence a raising of g.
What is the origin of g‐values diﬀerent from 2.0?

 For an electron that rotates into an empty orbital, the "current" created by the
rotating electron leads to a magnetic field that opposes the applied magnetic
field B0. This requires a larger applied field to achieve resonance, and hence a
lowering of g.
Mo(CN)8 3‐
7
EPR Spectra
Problems in observing EPR transitions:
H0
So, one must be careful to avoid “power saturation”:

i.e., driving the populations into exactly equal fractions
by driving transitions faster than spin relaxation
and therefore losing all net change in magnetization.
EPR Spectra
Problems in observing EPR transitions:
2. Zeeman transitions are magnetic dipole coupled only:

ms = ±1 therefore only small transition probability.
3. Transitions are often broad

relative to energy of transition.
Line broadening from site inhomogeneity,
leads to Gaussian shapes.
8
EPR Spectra
Solution to observing EPR transitions:
To enhance S/N and locate broad signals,

EPR spectrometers use modulation detection
 1st Derivative spectra: Signal Ampl / mag field
Instrumentally, the mag field is modulated using coils aligned with
mag field (Helmholtz coils) to oscillate H0 a bit (0.01 to 20 G)
at ~100 kHz. Detection is then phase locked to that frequency.
EPR Spectra typically given as 1st Derivative
9
EPR Spectra typically given as 1st Derivative
Absorption Derivative
 increasing g decreasing g
EPR QM
kinetic energy, e─ - nuclear attraction, e─ - e─

repulsion, nucl.-nucl. replusion, ligand field
“spin Hamiltonian”
Nuclear Zeeman (NMR)
Electronic Zeeman (EPR)
Zero Field Splitting (ZFS) from

spin-orbit coupling of angular mom.
Hyperfine Coupling of e─ & nuclear

moments (NMR & EPR)
10
g-value Anisotropy
g-value Anisotropy
Anisotropic Interactions always complicate things

(solids, frozen solutions, membranes etc.)
• with the applied field
• with surrounding magnetic nuclei
• among electron spins (if more than one, obviously)
Recall: Description of physical quantities

 Isotropic: scalars
 Directional: vectors
 Interactions between vectorial quantities: tensors
11
g is Anisotropic and Varies with Direction
g 'x'x' g 'x' y ' g 'x'z ' g xxx 0 0

Diagonalise
g ' y 'x' g ' y ' y ' g ' y 'z ' 0 g yyy 0
g 'z 'x' g 'z ' y ' g 'z 'z ' 0 0 g zzz
where g 'ij  g ' ji Principal values
Anisotropy: g  g zz  g iso
Asymmetry:   ( g zz  g xx ) g
For an arbitrary orientation

of a crystal in a magnetic field
In spherical coordinates:
g  ( g xx2 sin 2  cos 2 

 g yy
2
sin 2  sin 2 
 g zz2 cos 2  )1/ 2
12
g-value Anisotropy
Electronic Zeeman Hamiltonian (g is actually a tensor)
Uses & Applications of EPR
13
g-value Anisotropy
In a frozen solution, an ensemble of all possible orientations
of the metal site with respect to mag field vector.
A “powder spectrum” is then observed:
rhombic Powder axial Powder

(tetragonal)
absorbance absorbance
spectrum spectrum
g2 1st derivative 1st derivative

g3
g1
Often the g tensor has axial symmetry
Then: g║  g zz
g  g xx  g yy
┴
And: g  ( g 2 sin 2   g 2║ cos 2  )1/ 2
┴
14
Axial g-value Anisotropy
Absorption Derivative
Axial hyperfine (later): often a|| >> a┴ ~ 0

Axial g-value Anisotropy
15
Zero Field Splitting
Zero Field Splitting (also a tensor)
Zero Field Splitting (ZFS) vs. Zeeman Splitting

Kramer’s Doublets:
Effective half-integer spin
Zeeman states (S = 5/2, 3/2, 1/2,
Kramer’s Splittings etc.) that are split under
Doublets an applied magnetic field
to the ±Ms microstates
(Zeeman Splitting).
Example: A 4T1 state is split

under axial zero-field
splitting into three sets of
Kramer’s doublets (S =
5/2, 3/2, and 1/2), which
are separated by Zeeman
splitting to the ±3/2 and
±1/2 microstates.
© 2012, K.S. Suslick Zeeman axial rhombicity
16
Zero Field Splitting
ZFS from Ligand Field Distortions
17
FeIII, d5
Allowed Transitions for N nuclei with spins Ii

“Hyperfine” (HF) and “Superhyperfine” (SHF) are both due to
coupling between electron/paramagnetic spin and nuclear spins:
Hyperfine refers to interaction of a metal nucleus

with “its own” unpaired electron spin density.
Superhyperfine refers to interaction of unpaired electron spin density

with ligand nuclei.
For k sets of equivalent nuclei

with ni nuclei per set
with Ii nuclear spin per nuclei,
k
Total number of Hyperfine lines: N hf   ( 2ni I i  1)
k 1
18
Hyperfine Interactions
= interaction of paramagnetic spins with nuclear spins

For an isotropic coupling
between electron &
nucleus I = ½
Electronic Hyperfine
Zeeman (e- spin-nuclear spin)
Splittings Splittings
Isotropic Coupling between e- & nucleus I = ½
Recall:
S I
aiso/4
I
aiso/4
SI
S
S S
SI aiso/4
I
SI aiso/4
19
Hyperfine Interactions
Electronic Hyperfine
Zeeman (e- spin-nuclear spin)
Splittings Splittings
a = hyperfine
coupling
constant
Hyperfine with More than One Nucleus

1 spin ½ nucleus  (mI )   S  mI aiso
SI SI
3 eq. spin ½ nuclei
Sb Sb SI1I2I3 SI1I2I3
SI1I2I3 SI1I2I3
SI1I2I3 SI1I2I3
SI1I2I3 SI1I2I3
2 eq. spin ½ nuclei
SI1I2I3 SI1I2I3
SI1I2 SI1I2
SI1I2I3 SI1I2I3
SI1I2 SI1I2
SI1I2I3 SI1I2I3
SI1I2 SI1I2
SI1I2I3 SI1I2I3
SI1I2 SI1I2
20
k

k 1
e.g., benzene radical anion
1 set of 6 H (I=1/2)
Nhf = 2*6*½ +1 = 7 lines

k

k 1
e.g., [BH3●]— 1set of 3 H (I=1/2)

and 1 set of 1 11B (I=3/2)
Nhf = (2*3*½ +1)*(2*1*3/2+1) = 16 lines
EPR spectrum of [BH3●]— in solution.

The stick diagram marks the resonances
for the 11B(I=3/2) and the three protons.
(The remaining very weak resonances are

due to the radicals containing 10B (I=3).)
21
Oxidation of a Chromium(III) porphyrin to CrVPorph
(isotropic in solution)
S(Cr5+) = ½
I(14N) = 1
I(52Cr) = 0 (84% abundant)
Nhf = (2*4*1+1) = 9 lines
but also
I(53Cr) = 3/2 (9.6% abundant)
 (2*1*3/2+1) = 4 sets of 9 lines, weak
Two Spin Nuclei: Li+(13CO2─)

I(13C) = ½, I(7Li) = 3/2
12C

A(13C)>>A(7Li): Spin density mainly on 13C
22
Origins of Hyperfine Coupling
But Hyperfine can (is) also anisotropic
A  Aiso  Adipolar
Fermi contact Interaction Dipolar Interaction

Density of unpaired electron p,d,f orbital character
at nucleus (s-orbital character in SOMO.
in semi-occupied MO (SOMO). Averages out in soln.
ISOTROPIC, ANISOTROPIC
“Through-bond” “Through Space”
23
A Model Cu2+ system
Axial symmetry
I(65Cu) = 3/2 d9, S=1/2
g║ g┴
Axial hyperfine: often a|| >> a┴ ~ 0

24
HFS for Identification of Metal
Uses & Applications of EPR: Metal Identification
25
26
I = 3/2
27
28
Identification of Ligands: “Superhyperfine”
Identification of Ligands
29
Transition Metal EPR
• Complicated by the fact that transition metal systems
might have several unpaired electrons and several
approximately degenerate orbitals
• 3d elements important: only moderate spin-orbit coupling
• Ability to distinguish between high spin and low spin
complexes (in ligand fields): coordination number and
geometry accessible via EPR
• Difficult to observe EPR on systems w/ integer S systems
• Most common:
Ti3+(d1)S=1/2
Fe3+(d5) S=5/2 (high spin) often high anisotropy, S=1/2 (low spin)
Cu2+(d9) S=1/2 I=3/2 for 63Cu and 65Cu
Co2+(d7) S= 3/2 (high spin) S=1/2 (low spin)
ZFS in EPR
30
ZFS in EPR
Uses & Applications of EPR: Heme Proteins

Easy to tell L.S. Fe(III) from H.S. Fe(III)
31
powder spectrum for a rhombic g-tensor
Low spin Fe3+ in Powder

cytochrome P450 spectrum
1st derivative
s3
g xx g yy g zz
32
Slide 63
s3 There are many Cytochrome P450 enzymes, and they catalyse oxidation of various organic substrates
(e.g. medicines) in living organisms. At the centre of these enzymes is a haem centre (iron porphyrin),
bonded to the protein through an Fe-S bond to a cysteine side-chain. In the resting state of the
enzyme, the iron centre is in the +III oxidation state. Successive one-electron reduction, binding of
dioxygen, one-electron reduction, addition of two protons, and loss of water, leads to formal transfer of
an oxygen atom to the iron atom. The resulting key intermediate, referred to as "Compound I", formally
has an iron atom in the +V oxidation state (in fact, part of this charge is delocalised on the porphyrin
ring and the oxygen atom, so that the iron is better described as being in the +IV or even +III
oxidation state).
stuart, 2/22/2006
 Blumberg and Peisach’s plot of a large
family of low spin hemes showing how
data cluster into five domains. The first
four (C, B, H, O) have similar rhombic
distorHons but increasing axial field
strength. These assigned with histidine
(axial 5th site) but 6th ligand is methionine
C, neutral histidine H, anionic histidine B,
and oxide O. The fifth family (P) are the
P450 heme enzymes, with cysteine and
water as the axial ligands.
 The Blumberg analysis tends to work best
when V/Δ is quite large. These are known
as “Truth Tables”.
33
34
35
g values
36
37
Each nucleus experiences the hyperfine field

of only one electron.
Each (spin 1/2) nucleus then gives rise to two resonance

conditions depending on whether the electron hyperfine
field opposes or augments the applied field.
A strong radiofrequency (NMR) field induces NMR

transitions which are observed as a change in the
intensity of an electron resonance condition.
Electron Nuclear Double Resonance (ENDOR)
38
39
ENDOR
The ENDOR experiment (simplified)
NMR transition(3-4) at
1
Recall: I  a Thermal EPR 1-3
2
Equil. saturated.
4 | S I 
1 1 1
E4    s   I  a
2 2 4 1  1 
1 1 1
E3    s   I  a 3
|  S I  1  1
2 2 4
sat
| S I 
1 1 1
E2    s   I  a 2 1  1 
2 2 4
1 
1 1 1
E1    s   I  a 1
2 2 4 |  S I  1
40
ENDOR
 Relative populations are given by Boltzmann at thermal equilibrium
(I<<S, hence populations of 1 & 2, 3 & 4 assumed identical)
 Irradiate 1-3 transition (saturate at high power): that will give same
populations in 1 & 3.
 Irradiate system with RF (NMR) and sweep frequency while continually
saturating EPR transition;
observe the intensity of its absorption
 When RF frequency matches |I-a/2|, transition 3-4 will be induced,
restoring some population difference between levels 1&3
 More EPR absorption now possible: that is ENDOR signal
 Equally, when RF frequency matches |I + a/2| (1-4 transition), this time
a pumping from 1-4 occurs (as 4 has the higher population) and a
population difference between 1&3 is again achieved and EPR transition
enhance – the second ENDOR signal
 In practice, need to consider spin lattice relaxation processes
Orientation Selection
EPR
Hyperfine
couplings not
resolved
1H ENDOR
Toluene Solvent
Two wide doublets which give the hyperfine couplings to

protons in the C8H8 and C5H5 rings directly.
41
Electron Nuclear Double Resonance
Tetracene cations in sulphuric acid
EPR spectrum
ENDOR 1H (3 types of protons)
42

Electron Paramagnetic Resonance (EPR) Electron Spin Resonance (ESR)

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Electron Paramagnetic Resonance (EPR) Electron Spin Resonance (ESR)

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Electron Paramagnetic Resonance (EPR)

= Electron Spin Resonance (ESR)

I. Origins of EPR: Zeeman Interaction

II. Experimental considerations, Spectral presentation

III. Anisotropy and Spin-Orbit Coupling Components

IV. Hyperfine and Superhyperfine Splittings

V. Uses & Applications

Survey of Spectroscopic Techniques

© 2012, K.S. Suslick

© 2012, K.S. Suslick

EPR Pros & Cons

What EPR can tell us:

• Resolution is not as good as in NMR (broad features)

© 2012, K.S. Suslick

kinetic energy, e─ - nuclear attraction, e─ - e─

Nuclear Zeeman (NMR)

Electronic Zeeman (EPR)

Zero Field Splitting (ZFS) from

Hyperfine Coupling of e─ & nuclear

© 2012, K.S. Suslick

Zeeman Interactions: Energy Splittings

3. For one free electron: cf. EI = - mIħH0 (nucleus)

 B = Bohr magneton

5. Energy difference: gβ(+½)H - gβ(-½)H = gβH = h

 B = Bohr magneton

Zeeman Interactions: Energy Splittings for 1 e-

© 2012, K.S. Suslick

© 2012, K.S. Suslick

Zeeman Interactions: Energy Splittings

© 2012, K.S. Suslick

In the case of a free electron in

In the case of an e- in a molecule,

When unpaired electron couples to

h  (GHz ) 714.48  (GHz )

• For most organic radicals, g ≈ ge

Note for later: g is anisotropic and not a scalar but a tensor.

© 2012, K.S. Suslick

What is the origin of g‐values diﬀerent from 2.0?

© 2012, K.S. Suslick

So, one must be careful to avoid “power saturation”:

2. Zeeman transitions are magnetic dipole coupled only:

3. Transitions are often broad

© 2012, K.S. Suslick

To enhance S/N and locate broad signals,

© 2012, K.S. Suslick

EPR Spectra typically given as 1st Derivative

© 2012, K.S. Suslick

kinetic energy, e─ - nuclear attraction, e─ - e─

Nuclear Zeeman (NMR)

Electronic Zeeman (EPR)

Zero Field Splitting (ZFS) from

Hyperfine Coupling of e─ & nuclear

© 2012, K.S. Suslick

© 2012, K.S. Suslick

Anisotropic Interactions always complicate things

• with the applied field

• with surrounding magnetic nuclei

• among electron spins (if more than one, obviously)

Recall: Description of physical quantities

© 2012, K.S. Suslick

g 'x'x' g 'x' y ' g 'x'z ' g xxx 0 0

where g 'ij  g ' ji Principal values

© 2012, K.S. Suslick

Nhf = 26½ +1 = 7 lines

Nhf = (23½ +1)(21*3/2+1) = 16 lines