Nuclear magnetic resonance spectroscopy (NMR)
Proton Nuclear magnetic resonance ( 1H-NMR)
Principle: Range of NMR is (60-500MHz)
• When the nucleus of an atom is kept under strong
magnetic field nuclear energy levels are produced.
The nucleus is excited from its lowest energy
nuclear spin state the next higher one by
absorption of radio frequency.
Spin and the splitting of energy levels in a
magnetic field
• Subatomic particles (electrons, protons and
neutrons) can be imagined as spinning on their
axis.
• In most of the atoms these spins are paired against
each other and has no overall spin. However, in
some atoms the nucleus does possess an overall
The rules for determining the net spin of a nucleus
1. If mass number is even (mass number is the sum
of protons and neutrons)
a) If both number of protons and number of
neutrons are even. Protons and neutrons
separately pair-up inside nucleus, then the
nucleus has no net spin (I =0) such nuclei are not
NMR active. eg. 24He, 6C12 , 8O16
b) If both number of protons and number of
neutrons are odd (mass number will be even).
Protons and neutrons separately pair-up inside
nucleus leaving a half integral spin in each kind
of particles, they combine parallel give integral
spins for the nucleus (I =1,2,3...); such nuclei are
NMR active Eg. 1H2 , 7N14 I = 1
2. If mass number is odd
a) If mass number is odd, number of protons odd
and number of neutrons even or zero but, proton
pairing leaves odd half integral spins. Such
nuclei are NMR active eg. 1H1 = ½
b) If number of protons even and number of
neutrons odd. Net proton spin is zero but,
neutrons pairing leaves odd half integral spins
eg. 6C13 I= ½ such nuclei are also NMR active
Quantum mechanics tells us that a nucleus of spin
I=0 will have (2I+1) possible orientations. Nucleus
with spin ½ will have 2 possible orientations. In the
absence of an external magnetic field, these
orientations are of equal energy.
The precession of nucleus in a magnetic field
• Consider the behaviour of nucleus as a charged
particle in the magnetic field. Imagine a hydrogen
nucleus of spin +½ in a magnetic [Link]
nucleus is in the lower energy level (i.e. its
magnetic moment does not oppose the applied
field). The nucleus is spinning on its axis in the
presence of a magnetic field, this axis of rotation
will precess around the magnetic field
The frequency of precession is
termed as Larmor frequency. When
this frequency becomes identical to
the radio frequency, resonance
occurs and flipping of proton takes
place with the absorption of
radiation and spin orientation of
nucleus become -½ ie. The
nucleus "flips" the magnetic
moment so that it opposes the
applied field. Thus nucleus is in the
higher energy state.
The potential energy of the precessing nucleus is given
by E = - B cos
Where dipole moment of the nuclear magnet
B - applied magnetic field strength
angle between the direction of the applied field and the
axis of nuclear rotation.
The dipole moment of the nuclear magnet is =
constant is called the gyromagnetic ratio and is a
fundamental nuclear constant which has a different
value for every nucleus and h is the Planck's constant.
• Shielding and Deshielding: when a molecule is
placed in a magnetic field its electrons are caused
to circulate and thus they produce secondary
magnetic field. This induced magnetic field may
oppose or reinforce the applied field. Thus the two
possibilities are
• Shielding:
• Shielding: If the induced magnetic field opposes
the applied field, the net field felt by a hydrogen
nucleus in a molecule will be less than the applied
field, and it is said to be shielded. A more shielded
hydrogen nucleus absorb RF radiation at lower
frequency. Electron density around the nucleus,
greater will be the shielding effects. NMR signals
appear in the upfield region
• Deshielding: The induced magnetic field
reinforces the applied field, net field felt by a
proton in a molecule will be greater than the
applied field, and the proton is said to be de-
shielded. eg. deshielding is observed for protons
in an aromatic ring: A more deshielded proton
absorbs RF radiation at higher frequency
B
Chemical shift (
• Different protons in a molecule give signals at
different frequency. The difference in resonance
frequency of a nucleus that results from its
molecular environment is called its chemical shift.
It is measured with respect to a standard reference
TMS [tetramethylsilane (CH 3)4Si]
Chemical shift (=
• Problem: When 'H NMR of chloroform (CHCl 3),
is taken using a 100-MHz instrument and signal
appears at 728 Hz higher from the TMS signal.
Calculate the chemical shift.
Note: Resonance frequency of TMS is considered
as zero
Chemical shift = =
=7.28ppm
• The chemical shift may also be defined as the
shift in radio frequency from TMS due to
shielding or deshielding of hydrogen nucleus in
different structural environment.
Why TMS is chosen as a reference?
• In tetramethylsilane (CH 3)4Si, 12 protons are
identical and gives a single signal which appear at
lower frequency (upfield) due to lower
electronegativity of Silicon than carbon. The
protons of TMS are more shielded than most of
the organic compounds, So all the samples appear
at higher frequency (at downfield) than those of
the TMS. That is why it
is chosen as reference
Factors Affecting Chemical Shift
[Link]: Protons attached to more
electronegative atom shielding effect decreases
and chemical shift increases. In compounds of the
type CH 3 X, the shielding of the methyl protons
decreases as X becomes more electronegative.
2. Cumulative effects of electronegative
substituents: If the number of electronegative
substituents increases chemical shift also
increases.
Eg. the chemical shifts for various chlorinated
derivatives of methane
CHCl3 CH2Cl2 CH3Cl
chemical shift( 7.3 5.3 3.1
3. Distance from the electronegative atom: When
an electro negative atom is substituted in the
hydrocarbon chain, the electron density around the
protons de creases as the protons get closer
towards the electronegative atom. As the electron
density around proton decreases shielding effect
decreases value increases
e.g. 1-chloropropane CH3-CH2-CH2Cl
chemical shift( 1.0 1.4 3.4
4. Deshielding: In deshielding the induced
magnetic field reinforces the applied magnetic
field. Therefore deshielding effect increases the
delta For aromatic protons -value lies between
6.5 to 8.5.
5. Hydrogen bonding: Presence of hydrogen
bonding changes the values. The proton attached
with hydrogen bonding are deshielded and value
increases.
Interpretation of Chemical shifts in a molecule
1. The number of signals, tells us how many different
kinds of protons are present.
2. The position of the signals ( values) gives
information about the nature of protonic
environment.
3. The intensity of the signals is measured by the area
under each peak, which gives us the relative ratios of
the different kinds of protons.
For example CH3-O-CH2-CN
There are two kinds of protons, CH 3 protons and
CH2 proton. CH2 protons are less shielded (higher
value) due to electron withdrawals either side by
the electronegative atoms. The peak area ratio is
equal to 3:2 which proportional to number of H-
atoms in each set.
Spin-spin coupling: The splitting of NMR signals Every
hydrogen in a molecule spins and generates their own
magnetic field. The proton on neighbouring carbon will
generate magnetic fields whose magnetic moments will
interact with the magnetic moment of the external
magnetic field. This results in the splitting of the NMR
signal.
• The splitting of NMR signals due to coupling
interactions with neighbouring protons is called
spin-spin coupling or spin-spin splitting.
In a low resolution NMR spectroscopy give broad signal
but high resolution shows splitting of signal of proton.
General rules for spin-spin interaction
1. Chemically equivalent proton do not show
interaction, but environmentally different proton
show splitting.
2. Proton on adjacent carbon will normally couple
3. proton separated by four or more bonds will not
couple.
n+1 rule : This rule predict the splitting of signals
If ‘n’ protons are present in near by nucleus the peak
will split to n+1 lines.
Number of neighbouring Splitting (n+1)
proto n
0 0+1=1 Singlet
1 1+1=2 doublet
2 2+1=3 triplet
3 3+1=4 quartet
NMR spectrum of 1,1-dichloroethane(CH3CH-Cl2)
The CH3 protons split into a doublet under the influence of CH
protons and the peak intensity ratio will be 1:1
Similarly the CH protons split into a quartet with intensity ratio
[Link]. 1
The spins of three CH3 protons B 3
can align in four different ways 3
1
a doublet at 2.1 ppm for the methyl protons (CH 3 )
and a quartet at 5.9 ppm for the CH proton.
Magnetic Resonance Imaging
• Magnetic resonance imaging (MRI) is a medical
imaging technique used in radiology to visualize
internal structures of the body in detail.
• MRI use the property of (NMR) to image nuclei of
atoms inside the body.
• The body tissue contains lots of water, and the
protons ('H nuclei), which get aligned in a large
magnetic field.
• When a person is kept inside the powerful
magnetic field of the scanner, average magnetic
moment of many protons becomes aligned with
the direction of field.
• A suitable radio frequency (resonance frequency)
of current is turned on. The radio waves are
absorbed and flips the spins of the protons in the
magnetic field.
• Once the RF signal is removed the nuclei return to
equilibrium is referred to as relaxation.
• During relaxation, the nuclei lose energy by
emitting their own RF signal.
• This signal is called free-induction decay (FID)
response signal and is measured by a conductive
field coil placed around the object being imaged.
• This images are reconstructed to obtain 3D grey-
scale MRI images.
• Relaxation times for molecules to regain their
natural alignment vary depending on the type of
tissue being scanned.
• Computer will record the amount of time taken
by the molecule to realign themselves.
• This allows the scan to detect different types of
tissue as it make up the body.
Applications of MRI
• Diffusion MRI: It uses the diffusion of water
molecules in biological systems. Useful for
diagnosis of neurological disorder and help in
better understanding of connectivity of central
nervous system.
• Magnetic resonance spectroscopy: Used to
measure the levels of different metabolites in
body the particularly in brain
• Magnetic resonance angiography: It generates
the picture of the arteries
• Functional MRI: It measures that change in
brain that are due to changing neural activity