Electromagnetic radiation spectrum

NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY

PRINCIPLE AND APPLICATION IN STRUCTURE ELUCIDATION

Professor S. SANKARARAMAN
Department of Chemistry
Indian Institute of Technology Madras
Chennai 600 036

sanka@iitm.ac.in

HISTORICAL PERSPECTIVE
Discovery of NMR phenomenon in 1945
Purcell, Torrey and Pound Harvard University USA
Bloch, Hansen and Packard Stanford University USA
When ethanol was placed between pole pieces of an
electromagnet and irradiated with electromagnetic
radiation it absorbed radiation in the radio frequency
region. When the magnetic field was turned off no
absorption was observed.
Purcell and Bloch Nobel prize in Physics 1952
For the discovery of NMR.

The first published high resolution NMR spectrum of ethanol at 30 MHz

F. Bloch, W. W. Hansen, M. E. Packard, Phys. Rev. 1946, 69, 127

NMR magnetic properties of atomic nuclei

Atomic nucleus has mass and it spins on its own axis
Due to the spin, it possesses angular momentum (P)
Due to the charge and the spin it possesses magnetic momentum (m)
Only certain nuclei have non-zero magnetic moment. In others the
Net magnetic momentum can be zero
Only nuclei with non-zero magnetic moment are magnetically active
Both (P) and (m) are vector quantities and also quantized
The ratio of magnetic momentum to angular momentum is called
Gyromagnetic ratio. It is very characteristic of a given nuclei.

Gyromagnetic ration = [g] = (m)/(P)

Basic theory of NMR Spectroscopy

Nucleus should be magnetically active
non-zero magnetic momentum
According to quantum mechanics angular momentum
can have only certain fixed values (eigen states)

P = (m)(h/2p) where m is the magnetic quantum number of the

nucleus
In the presence of an external magnetic field(m) can have
(2I+1) values, namely (+I), (I-1), (I-2)..(-I)
where (I) is the spin quantum number of the nucleus
For I = , two states are possible (+) and (-)
For I = 1, three states are possible (+1, 0, -1)
For I = 3/2 , four states are possible (3/2, +1/2, -1/2, -3/2)

Nuclear Spin (I):

A simple way to find out nuclear spin

Atomic mass
Even
Even
Odd

Atomic number
Even
Odd
Even or Odd

Spin
zero
multiple of 1
multiple of

Examples:

I=0
I = integer
I = half integer

12C , 16O
6
8
14N (1), 10B (3), 2H (1)
7
5
1
1H (1/2), 13C (1/2), 15N
1
6
7
17O (5/2) 33S (3/2)
8
16

(1/2)

Properties of some common NMR nuclei:

Nucleus

Spin

g
(rad T-1 s-1)

1H

Natural
abundance (%)

26.7

99.9

4.107

0.015

13C
6

6.72

1.10

19F
9

25.18

100

31P

10.84

100

-5.32

4.67

2H

15

29Si

14

NMR of spin nuclei, namely proton and carbon-13

In the absence of external magnetic field the magnetic moment
vectors will be randomly oriented
In the presence of applied external magnetic field (Bo) two
orientations are possible, namely (+1/2) and (-1/2)
The two orientations, one aligning with external field (-1/2)
and another opposing the external field (+1/2),
differ in energy.
The energy difference depends on the strength of applied
magnetic field

Nuclear spins in an external magnetic field for I = 1/2

+ 1/2 b

E = hn = (h/2p)Bog
E = hn
no field

Bo
increasing filed strength Bo

n = (Bog)/2p

- 1/2 a

Distribution of nuclear spins:

Na / Nb = exp(-DE/kT)
Bo (T)

n (MHz)

DE (J)

Na / Nb

T oC

2.35

100

6.7 x 10-26

17 ppm

17

4.70

200

22.5 x 10-26

57 ppm

17

7.0

300

33.5 x 10-26

85 ppm

17

2.35

100

6.7 x 10-26

28
13

-100
+100

Higher the magnetic field strength

higher the sensitivity and resolution
Lower the temperature
higher the sensitivity
A 400 MHz NMR instrument is more sensitive
as well as more resolving than a 60 MHz NMR
instrument

The energy gap between the spin states corresponds

to radio frequency region
Application of radio frequency causes the absorption
of the same due to excitation of nuclear spins from
lower energy level to upper energy level when the two
energies match (resonance condition)
The spins in the excited state return back to ground
State by (a) spin lattice relaxation and (b) spin-spin
relaxation

Concept of chemical shift:

n = (Bo g)/ 2p
Nucleus is surrounded by electrons
Electrons have charge as well as spin
The magnetic field due to the spinning electron
shields the nucleus from the external magnetic
field. This is diamagnetic shielding.
The nucleus does not feel Bo, but Beff = Bo(1-s)
Beff is the effective magnetic field felt by nucleus
s is the shielding constant

s - the shielding constant

Characteristic of the chemical environment
of the proton
Depends on the electron density around the protons

n = (Beff g) / 2p = [Bo(1-s) g / 2p]

Since s is different for chemically different protons
the resonance frequency of chemically different
protons will be different chemical shift

Definition of chemical shift d:

It is inconvenient to refer to proton frequency as
398.432 MHz
Instead of actual frequencies of resonances, a reference
is taken and the frequencies are calibrated with
respect to the reference
d (in Hz) = n sample n reference (Spectrometer dependent)

d=

(n sample n reference) x 106

spectrometer frequency

(in ppm)

Chemical shift expressed in d is a dimensionless quantity and

also does not depend on the spectrometer frequency

Reference for 1H-NMR spectroscopy:

Tetramethylsilane (TMS) is used as a reference

The chemical shift of TMS is lower than most protons in
organic molecules, so it is taken as zero
All the protons in TMS are equivalent and hence only one
signal for all the 12 protons high signal intensity
TMS is a liquid and miscible with most solvents
It is also volatile and hence easy to remove
It is inert and does not react with the samples

Chemical shifts and scan widths

10 ppm
600 Hz

0
0

60 MHz spectrometer
10 ppm

4000 Hz

0
400 MHz spectrometer

d 2 ppm in a 60 MHz spectrometer is 120 Hz

d 2 ppm in a 400 MHz spectrometer is 800 Hz

Factors affecting chemical shift:

1. Electronegativity, inductive and resonance effects
TMS = 0.0

CH4 = 0.23 (all in ppm)

MeI
MeBr
MeCl
MeF

2.2
2.6
3.1
4.3

MeCl
CH2Cl2
CHCl3

3.1
5.3
7.2

MeOH
MeF
MeNO2

3.4
4.3
4.3

2. Anisotropic effects:
Spherical electron density induced magnetic field
will be uniform in space isotropic effect
For example s electron spherical

Non-spherical electron density induced magnetic

field will be non-uniform in space anisotropic
Example: p electron cloud of aromatic ring, C=C
and C=O type most common feature of organic
molecules

Diamagnetic anisotropy in ethylene and acetylene

H d 1.80
+

d 5.28

+
H

deshielding

- deshielding

+
shielding

shielding

Bo

Bo

Diamagnetic anisotropy in aromatic ring

Ring current effect
+

shielding

+
Bo

Bo

d 7.28
H
deshielding

Diamagnetic anisotropy in carbonyl group

d 9.5

+
shielding

Bo

deshielding

Examples of effect of anisotropy on chemical shift

1.32

1.03
H

H -0.51

1.27
Me

0.70
Me

H
8.14-8.67

H
H

7.29
H

1.44

-4.2

H
H

H
H

2.82
H

H 9.3

-3.0

Anisotropic effect of sigma bond

1.12
H
H
1.62

4.23
H

NO2
NO2

H
4.43

3.37
H

OH
OH

H
3.93

CH3 CH2 - OH

OCH2Ph
O
PhCH2O
PhCH2O
PhCH2O

SPh

Solvent effect on chemical shift: (a) in CDCl3, (b) Benzene-d6

Effect of neighboring protons spin-spin coupling

a
Absence of any interacting protons
d
No neighboring protons
No spin-spin coupling only a single peak for each
chemically different proton

Consider two protons Ha and Hb - neighbors

Case 1
Case 1
no spin-spin interaction

Case 2
spin-spin interaction

bb
da

ba

db

Case 2

ab

Jab

Jab

da

db

aa
Ha Hb
only two transitions
one for Ha and one
for Hb

four transitions,
two each for Ha and
Hb

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/nmr/nmr1.htm

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/nmr/nmr1.htm

spin-spin coupling constant depends on

1. Distance between the coupling partners intervening bonds

2. Dihedral angle between the coupling partners for vicinal protons
3. Coupling constant is largest when dihedral angle is 180o
and very small when dihedral angle is 90o
4. In freely rotating bonds (like in alkyl chains) average

J values are obtained

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/nmr/nmr2.htm#pulse

Spin spin splitting patterns for I = nucleus like 1H:

The no. of lines from coupling = (2nI+1) = (n+1) for I =
Where n is the number of equivalent protons that couple

Ha

C C

Hb

Ha and Hb - each a doublet with Jab

Ha

C C Hb

Ha - triplet and Hb - doublet with Jab

Hb
Ha

C C

Hb Ha and Hb - each triplet with Jab

Ha Hb
CH3 - CH2

CH3 - triplet and CH2 - quartet

CH3-CH2-CH2

CH3 - triplet, CH2 - sextet, CH2 - triplet

CH3-CH-CH3

CH3 - doublet, CH-septet

Line intensities of multiplets for spin nucleus

Corresponds to the coefficients of binomial expansion
Can be obtained simply from Pascals triangle

1
11
121
1331
14641
1 5 10 10 5 1

1H-NMR chemical shifts of various types of protons

http://www.cem.msu.edu/~reusch/VirtualText/Spectrpy/nmr/nmr1.htm

Magnitude of some coupling constants J in Hz

Chemically equivalent protons

protons having same chemical environment
hence the same chemical shift (d)
Homo and enantiotopic hydrogens in organic
molecules
isochronous
Magnetically nonequivalent protons
protons having different magnetic environment
different coupling constant with other protons
diastereotopic hydrogens in organic molecules
For two protons to show spin-spin coupling they have to be
magnetically non-equivalent. They may or may not be chemically
non-equivalent

Parameters obtainable from a 1H-NMR spectrum:

1. Chemical shift values of various protons

2. Coupling constant values from multiplets
3. Relative ratio of signal intensities area under the peaks
proportional to the number of protons responsible for
each signal
4. Relative mole ratios of components in a mixture

CH3CH2Br

CH3COOCH2C6H5

HCOOCH2CH3

A
C4H8O

CH3COCH2CH3

B
C4H8O2
CH3COOCH2CH3

C5H7NO2

CH3CH2O

CH2C N

C4H7O2Br

CH3-CH2-CH(Br)-COOH

H3C

CH3

CH2CH3

CH3
H3C
CH3

CH3
H
CH3

OH O
H

Ha
O
CH3
Hb

Ha and Hb are diastereotopic due to the adjacent chiral center

CH3

H
H3C
H

H
O
O
CH3

CH3

H3C

CH3
O

H
H

CH3CH2 O
O

O
O CH2CH3

O
O
O
O

CH2CH3
CH2CH3

H3C

C10H12O3

H
O

CH3

3H
3H
5H

1H

CH2CH2OH

2H
5H

2H

1H

C9H11Br

CH2CH2CH2Br

5H
2H

2H
2H

NH2

C6H5NBr2

Br

Br

H
H

2H

1H

2H

Second order spectra:

When the chemical shift values are very close and
the difference in chemical shifts are comparable to J values
one finds second order effects in the NMR spectra
Typically when (Dd)/J is less than 10, second order effects
are seen in the spectra
Unusual intensity of multiplets
More than expected number of lines in multiplets
are characteristic features of a second order spectrum

Examples of spin systems that show second order effects:

X
R

Ha

Y
X

Z
R

Hb

Ha
Hb

mutually coupled ab spin system

da, db, Jab

Ha

Hx

mutually coupled amx spin system

Hm

da, dm, dx, Jam, Jax, Jmx

X
Ha

Ha'

Hb

Hb'
Y

Ha'

Ha
X

Hb

Y
Hb'

Note: Ha and Ha' are chemically

equivalent but magnetically
non-equivalent protons. similarly
Hb and Hb' are.

mutually coupled aa'bb' spin system

da, db, six different coupling constants

C6H8N2
H3C

2H

1H

1H

1H

NH2

3H

Dimethyl cyclopropanedicarboxylate

Variable Temperature NMR Spectroscopy

Study of dynamic properties of molecules

Conformational changes
Restricted rotation around C-C and C-X bonds
Aggregation phenomena
Fluxional properties of molecules
Temperature range typically -150 oC to +150 oC
provided solubility and solvent mp/bp permit
Processes with activation barrier typically in the range
of 8-25 Kcal/mole can be studied

Restricted rotation around C-N bond

In amides
O
Me

N
Me
O

O
Me
N
Me

Me
N
Me

Activation barrier = 22 Kcal / mole

Conformational changes

Interconversion of chair
form of cyclohexane

H
H

Activation barrier = 10 Kcal / mole

Cyclohexane-d11

D2

D2

H
NH

D2

D2

NH

Fluxional behaviour of
large annulenes
H

H
H

H
H
H

H
H

H
H

H
H

[18]annulene

Bullvalene most fluxional molecule

Synthesis: Schrder 1963
C10H10 isomer
C3v symmetry
Degenerate Cope rearrangement [3,3]-shifts
Activation barrier = 11.8 Kcal/mole

H
H

H
H

H
H

H
H

Me
2
Al
Me
Me

Me

Me
Al
Me

Me
Al

Me

Me

13C-NMR

spectroscopy

Carbon-13 nucleus is spin active wih I =

Hence C-13 NMR spectroscopy is possible and very useful
in organic structure elucidation

Abundance of Carbon-13 is very low, only about 1.1%

The gyromagnetic ratio of Carbon-13 is also low, it is about
1/4th of proton gyromagentic ratio
Both these factors are responsible for the poor sensitivity of
Carbon-13 NMR spectroscopy, it is about 1/(6400)th of
proton

FT-NMR technique
Two types of NMR spectrometers
Continuous wave (CW)
Fourier Transform (FT)

In CW spectrometer either the magnetic field or the

radio frequency is swept, bringing each nuclei to
resonance one at a time signals are recorded one at a
time hence very time consuming because each SCAN
has to be accumulated and averaged
In FT technique a short pulse of radio frequency is
applied that bring all the nuclei to resonance. The nuclei
are allowed to relax to ground state and the resulting
free induction decay is FOURIER transformed

FREE INDUCTION DECAY OF EXCITED NUCLEI - FID

Signal
intenisty

The concept of time domain-frequency domain spectroscopy FT method

Effect of signal averaging on

S/N ratio

Carbon-13 NMR is usually recorded under conditions

of proton decoupling
That is all the C13-H1 coupling are decoupled by
irradiation (saturation) of all the protons
Therefore only a single signal is observed for each of
the chemically different carbons
From the symmetry of the structure one can easily
predict the number of signals expected for a compound

Groups like acetylene carbon, CN, CO, quaternary carbon

(no protons) are easily detected by Carbon-13 NMR

Broad band decoupling: Carbon-13 spectra are recorded with

simultanous saturation of the proton spins using a second radio
frequency corresponding to the protons. This results in complete
decoupling of the protons and only carbon peaks are seen in the spectrum.

Gated coupling: Decoupler is switched on only during the delay time

and it is off during the data acquisition.
NOE enhancement is retained and the Carbon-13 spectrum is
proton coupled
Off-resonance decoupling: Decoupling with the radio frequency
that is not exactly that of protons but few hundred hertz displaced.
Splitting only due to the protons directly attached to a carbon are seen.
CH3 quartet, CH2 triplet, CH doublet, Quaternary C - singlet

Unlike proton NMR the signal intensities of Carbon-13 spectrum is

usually not quantitative.
1. The relaxation times of Carbon-13 nuclei are much longer than that of
protons.
2. Nuclear Overhauser Effect (NOE).
Relaxation mechanisms:
Spin-Lattice or Longitudinal Relaxation: (T1)
Relaxation by dispersing energy to the surroundings (lattice).
Spin-Spin or Transverse Relaxation: (T2)
Relaxation by dispersing energy to other spin active nucleus.

Nuclear Overhauser Effect:

Enhancement of signal intensities due to heteronuclear coupling

For example: Carbon-13 signal intensities are enhanced due to

irradiation (decoupling) of the protons.
The major relaxation route for Carbon-13 nucleus involves
dipolar transfer of its excitation energy to protons that are
directly attached to it (transverse relaxation).
NOE effect will be maximum for CH3, CH2, CH and none for
Quaternary carbon.
Therefore peak intensities of CH3 > CH2 > CH > C

Relaxation agents:
Paramagnetic relaxation agents such as Cr(acac)3 reduces
the longitudinal relaxation
This allows faster signal averaging

10-100 mM relaxation agent is needed the solution takes on

a slight pink-purple hue
Net result: Signal intensities of quaternary carbons are
enhanced

(a) Spectrum of camphor without relaxation agent

(b) With relaxation agent, Cr(acac)3 added, under otherwise identical conditions

Chemical shift range in 13C-NMR spectrosocpy

CH3CH2 O
O

2
1

O CH2CH3
4

2,3

4
1

O
O
1 O

4
3

CH2CH3
CH2CH3
5

3
CDCl3
2
1

H3C

NH2

O
CH3CH2O

CH2C N

Distorsionless enhancement by polarization transfer (DEPT)

2D Correlation Spectroscopy (COSY)

HO

OH
N3

O
O
O

H-H COSY of 2-chlorobutane

AcO

OAc

N3

AcO

OAc

H
N

O
O

O
A

O
B

N
N

N
N

HMQC NMR of menthol

Het-2D-J resolved
2-chlorobutane

a-Pinene

References:
J. B. Lambert, H. F. Shurvell, L. Verbit, R. G. Cooks, G. H. Stout
Organic Structural Analysis
Macmillan, New York, 1976
W. Kemp
Organic Spectroscopy, 3rd Edition, Macmillan, New York, 1991

D. H. Williams, I. Fleming
Spectroscopic Methods in Organic Chemistry, 4th Edition
Tata McGraw Hill, New Delhi, 1988
R. M. Silverstein, G. C. Bassler, T. C. Morrill
Spectroscopic Identification of Organic Compounds, 5th Edition
John Wiley, New York, 1991
H. Gnther
NMR Spectroscopy, 2nd Edition
John Wiley, New York, 1994

John Wiley, 1998

Timothy D. W. Claridge, High-Resolution NMR Techniques in Chemistry, Pergamon, 1999