NUCLEAR MAGNETIC

RESONANCE

A branch of spectroscopy where the spin

changes at the nuclear level when a
radiofrequency energy is absorbed in the
presence of magnetic field is studied.

It uses radio-frequency waves to induce transitions between

nuclear energy states in a magnetic field.
 Principle of NMR:
A nucleus with odd mass no. or an
odd atomic no. has a nuclear spin
which generates a magnetic field
along the axis of spin.
In the absence of external magnetic
field the nuclei are randomly
oriented. When the external field is
applied, nuclei align themselves
parallel to external magnetic field.
When energy is applied in the form
of radio-frequency wave transition
of proton from lower energy to
higher energy state occurs and nmr
signal is thus recorded.
 H1 - NMR
When Proton(hydrogen) is studied, then it is called as PROTON
MAGNETIC RESONANCE(PMR).
Any proton/nuclues with odd mass number spins on its own
axis.when external magnetic field is applied, nuclues spins and c\a
magnetic moment is created giving rise to frequency called
precisional frequency.
when Applied frequency = precisional frequency, nmr signal is
recorded.
Transition of proton from lower energy to higher energy takes
place.
 NUCLEAR & ELECTRON SPIN

All nuclei are assigned a spin quantum number, I, on the basis of

thenumber of protons and neutrons in the nucleus, the values being
zero,half-integral of integral. The number of permitted orientations in
Space.
hence quantized energy levels, or spin states, that can be adopted
by
nucleus subjected to an externally applied magnetic field is given as
(2I + 1). Thus, for I = 12, two orientations,
or energy levels, are possible. The following should be noted.
A spinning nucleus, being charged, generates a magnetic moment
vector, or dipole, along its axis of rotation.
The NMR Spectrometer
 Basic Theory NMR
Only nuclei with spin number (I) 0 can absorb/emit
electromagnetic radiation.

Even atomic mass & number I = 0 (12C, 16O)

Even atomic mass & odd number I = whole integer
(14N, 2H, 10B)
Odd atomic mass I = half integer (1H, 13C, 15N, 31P)

The spin states of the nucleus (m) are quantized:

Properly, m is called the magnetic quantum number

 Basic Theory NMR

For 1H, 13C, 15N, 31P (biologically relevant nuclei with I = 1/2):

magnetic moment (m),

It is a vector quantity that gives the direction and magnitude

(or strength) of the nuclear magnet
h is the Planck constant
g is the gyromagnetic ratio, and it depends on the nature of each
nuclei.
.
The energy of a spin in a magnetic field will depend on the
magnetic field, which we call Bo, and m:
 Basic Theory NMR
When the Bo field is applied, spins have to possible energy limits. In
one we are in favor of the field, and in the other one we are
against it. The energy is the dot product of the corresponding
vectors:
ENERGY, EE==-- . . BBo
o

Population ratio

E/ RT
NN //NN ==ee E/ RT

 Basic Theory NMR
Net magnetization
If we decompose the vectors in z and <xy>, we get:
z

Well have a slight excess of spins

x aligned with Bo, but at any angle
y with respect to z. The distribution
Bo is proportional to N / N .

z z

z
=
Mo
x
y
y
Bo
x = 0
The net magnetization is aligned with Bo,
and this is what we use in NMR.
 Chemical Shift

Measured in parts per million.

Ratio of shift downfield from TMS (Hz) to total
spectrometer frequency (Hz).
Same value for 60, 100, or 300 MHz
machine.
Called the delta scale.
Frequency of signal frequency of reference x 106
Chemical shift, =
Spectrometer frequency
Protons in a Molecule

Shielding effect

e.g. CH3 - CH2 - CH2 - OH

0.92 1.57 3.58
 CHEMICAL SHIFT:
Depends on carbon group attached:

onacarbonyl (aldehyde):
O
9- 10ppm
C H
tocarbonyl (aldehydes&ketones):
O C/H
C C H 1.6- 3ppm
C/H
toC=C(allylic):
C/H
C C C H 1.6- 3ppm
C/H
toaromaticring(benzylic):
C/H
C H 1.6- 3ppm
C/H
 Depends in hybridization
C-C: C/H
means must
be C or H, not
C/H C H 0.2 to 2 ppm
O, N, or X C/H

C=C: C/H
C C H 4.5 to 7 ppm

=
C=C:
C C H 1.6 to 3 ppm

Aromatic: H 6.8 to 8 ppm

 Depends on anisotropy
Acetylenic Protons, 2.5

=>
 NMR Signals

The number of signals shows how many

different kinds of protons are present.
The location of the signals shows how
shielded or deshielded the proton is.
The intensity of the signal shows the
number of protons of that type.
Signal splitting shows the number of
protons on adjacent atoms.
 NMR Spectra

Carboxylic Acid Proton, 10+

 Intensity of Signals

The area under each peak is proportional to the

number of protons.
Shown by integral trace.
 EG:1,1,2-Tribromoethane
Nonequivalent protons on adjacent carbons.

=>
 Spin-Spin Splitting

Triplet: 2 Adjacent Protons

=>
 Spin-Spin Splitting
The N + 1 Rule

If a signal is split by N equivalent protons,

it is split into N + 1 peaks.
 Coupling Constants

Distance between the peaks of multiplet

Measured in Hz
Not dependent on strength of the external
field
Multiplets with the same coupling
constants may come from adjacent groups
of protons that split each other.
 Coupling Constants
Complex Splitting
a c
H H
C C
Hb

Signals may be split by adjacent protons, different

from each other, with different coupling constants.
Example: Ha of styrene which is split by an
adjacent H trans to it (J = 17 Hz) and an adjacent H
cis to it (J = 11 Hz).
 Carbon-13 nmr:

C has no magnetic spin.

12

13C has a magnetic spin, but is only 1% of

the carbon in a sample.
The gyromagnetic ratio of 13C is one-
fourth of that of 1H.
Signals are weak, getting lost in noise.
Hundreds of spectra are taken, averaged.
Combined 13 C
and 1H Spectra

=>
 Fourier Transform NMR
Nuclei in a magnetic field are given a radio-frequency pulse close
to their resonance frequency.
The nuclei absorb energy and precess (spin) like little tops.
A complex signal is produced, then decays as the nuclei lose
energy.
Free induction decay is converted to spectrum.

FT

FT

FT
Basics of 2D NMR Experiment
General Scheme for 2D NMR
COSY Spectrum

H-3 > H-5 > H-10 > OH

H-10 -> H-9
H-3 > H-16
H-16 > H-11
2D NMR
 WHY 2D NMR?
As you can see, 1D protein spectra are far too
complex for interpretation as most of the signals
overlap heavily. By the introduction of additional
spectral dimensions these spectra are simplified
and some extra information is obtained. The
invention of multidimensional spectra was the
major leap in NMR spectroscopy apart from the
introduction of FT-NMR. Consequently, both
techniques were acknowledged by a nobel prize.
Anatomy of a 2D experiment:
 2D NMR:
The construction of a 2D experiment is simple: In addition to preparation and detection
which are already known from 1D experiments the 2D experiment has an indirect
evolution time t1 and a mixing sequence. This scheme can be viewed as:

Do something with the nulcei (preparation),

let them precess freely (evolution),
do something else (mixing),
and detect the result (detection, of course).
After preparation the spins can precess freely for a given time t1. During this time the
magnetization is labelled with the chemical shift of the first nucleus. During the mixing
time magnetization is then transferred from the first nucleus to a second one. Mixing
sequences utilize two mechanisms for magnetization transfer: scalar coupling or dipolar
interaction (NOE). Data are acquired at the end of the experiment (detection, often called
direct evolution time); during this time the magnetization is labelled with the chemical
shift of the second nucleus.
 NMR APPLICATIONS:
Structural (chemical) elucidation
Natural product chemistry.
Organic chemistry.
Analytical tool of choice for synthetic chemists.
Study of dynamic processes

Reaction kinetics.
Study of equilibrium (chemical or structural).
Structural (three-dimensional) studies
Proteins.
DNA/RNA. Protein complexes with DNA/RNA.
Polysaccharides
Drug design

Structure Activity Relationships (SAR) by NMR

Medicine - Magnetic Resonance Imaging (MRI)

THANKS FOR BEING SUCH A
CAPTIVE AUDIENCE!