Nuclear Magnetic Resonance (NMR)

Lecture by :-
Mohit Ananda
Assistant Professor
 Introduction
 Nuclear Magnetic Resonance (NMR) is a spectroscopic method in which
many nuclei may be studied , but hydrogen and carbon are most commonly
available, while other nuclei such as fluorine – 19, phosphorus -31, nitrogen
– 14, nitrogen – 15, oxygen-17 can be studied selectively.
 Felix Bloch and Edward M Purcell shared the noble prize in physics for
their discovery of NMR phenomenon in simple nucleus of the hydrogen
atom .
 NMR spectroscopy is a powerful analytical technique used to characterize
organic molecules by identifying carbon-hydrogen frameworks within
molecules.
 Two common types of NMR spectroscopy are used to characterize organic
structure: 1H NMR is used to determine the type and number of H atoms in
a molecule; 13C NMR is used to determine the type of carbon atoms in the
molecule.
 The source of energy in NMR is radio waves which have long wavelengths,
and thus low energy and frequency. When low-energy radio waves interact
with a molecule, they can change the nuclear spins of some elements,
including 1H and 13C.
 WHAT IS A SPIN
 A rotating object possess a quantity called angular momentum given by the right
hand thumb rule.
 Spin is a type of angular momentum that does not vanish when at absolute zero! So
a physical motion to represent this type of angular momentum is not with out error.
 Spin is a fundamental property of electron and nucleus like mass, electric charge
and magnetism.
 NUCLEAR SPIN
 The nuclei of some atoms have a property called “SPIN”.
 Elements with either odd mass or odd atomic number OR both
have the property of nuclear “spin”. (Quantized angular
momentum and a magnetic moment)
 Each spin-active nucleus has a number of spins defined by its
spin quantum number, I.
 Many (but not all) nuclei have a spin (I). Quantum mechanically I
(spin quantum number) can have 2I+1 orientations in an external
magnetic field B with integral differences ranging from +I to – I.
 Atoms with odd mass number have spin I as integer multiples of one half (1/2, 3/2,
5/2 )
 Atoms with even mass number but odd numbers of protons and neutrons have spin I
as integer numbers (1, 3, 5, 7 …)
 Atoms with even mass number and even atomic number have zero spin.
 I = 2 does not exist
 I > ½ are known as quadrupolar nuclei.
 In the absence of an applied magnetic field, all the spin states of a given nucleus are of
equivalent energy, and in a collection of atoms, all of the spin states should be almost
equally populated with the same number of atoms having each of the allowed spins.
 When a charged particle such as a proton spins on its axis, it creates a magnetic field. Thus,
the nucleus can be considered to be a tiny bar magnet.
 Normally, these tiny bar magnets are randomly oriented in space. However, in the presence
of a magnetic field B0 , they are oriented with or against this applied field. More nuclei are
oriented with the applied field because this arrangement is lower in energy.
 The energy difference between these two states is very small (<0.1 cal).
 When external magnetic field is applied to the nucleus, protons have their magnetic
moment either aligned with the field or opposed to it.
 In a magnetic field, there are now two energy states for a proton: a lower energy
state with the nucleus aligned in the same direction as B0, and a higher energy
state in which the nucleus aligned against B0.

 When an external energy source (hn) that matches the energy difference (DE)
between these two states is applied, energy is absorbed, causing the nucleus to
“spin flip” from one orientation to another.

 The energy difference between these two nuclear spin states corresponds to the
low frequency RF region of the electromagnetic spectrum.
PRECESSIONAL MOTION (orbit)
ABSORPTION OF ENERGY
 The energy absorption is a quantized process and the energy absorbed must equal the energy
difference between the two states involved.

 The energy difference between the ground and excited states may be expressed by the
equation
δE = γhB0
γ = magnetogyric ratio = μ/Iħ (proportionality constant)
Μ = maximum observable component of the magnetic moment
 A spinning nucleus in a magnetic field (B0 ) experiences B0 as a torque about an axis
perpendicular to the axis of rotation.
 This causes the spin axis to precess about the direction of the field.
 An increase in the strength of B0 does not produce an energy transition of the nucleus,
but simply increases the precessional frequency.
 For a given value of B0 the precessional or larmor frequency is given by the equation
υ0 =(γ/2π) B0
Introducing Planck's constant (h) on both sides
h υ0 =(γh/2π) B0 = γh B0 = δE
 If the nucleus is irradiated with radio frequency energy where
 h υ0 = γh B0 = δE then, when it is in the ground state, it can absorb radiation energy.
 Radio frequency equal to the precessional frequency of the nucleus will
initiate resonance in the precessing spin axis and cause the nucleus to flip
into a higher energy state.

 The radiation energy required for such a transition falls within the
radiofrequency region of the electromagnetic spectrum and, in an NMR
experiment, is applied to the same at right angles to the magnetic field.
 Figure shows the origin of nuclear magnetic “resonance”
Theory of NMR spectra
 Protons in different environments absorb at slightly different frequencies,
so they are distinguishable by NMR.
 The frequency at which a particular proton absorbs is determined by its
electronic environment.
 The size of the magnetic field generated by the electrons around a proton
determines where it absorbs.
 NMR absorptions generally appear as sharp peaks.
 Increasing chemical shift is plotted from left to right.
 Most protons absorb between 0-10 ppm.
 The terms “up field” and “downfield” describe the relative location of
peaks. Up field means to the right. Downfield means to the left.
 NMR absorptions are measured relative to the position of a reference peak at 0 ppm
on the scale due to tetramethylsilane (TMS). TMS is a volatile inert compound that
gives a single peak upfield from typical NMR absorptions.
 Nuclei are surrounded by electrons.
 The strong applied magnetic field (Bo ) induces the electrons to circulate around
the nucleus (left hand rule). This circulation is called as local diamagnetic current,
generates a counter magnetic field which opposes the applied magnetic field.
 This phenomenon is called as diamagnetic shielding or magnetic anisotropy
 The induced circulation of electrons sets up a secondary (induced) magnetic
field (Bi ) that opposes the applied field (Bo ) at the nucleus (right hand
rule).

 We say that nuclei are shielded from the full applied magnetic field by the
surrounding electrons because the secondary field diminishes the field at the
nuclei.
 The electron density surrounding a given nucleus depends on the electronegativity
of the attached atoms.
 The more electronegative the attached atoms, the less the electron density around
the nucleus in question.
 We say that that nucleus is less shielded, or is deshielded by the electronegative
atoms.
 Deshielding effects are generally additive. That is, two highly electronegative
atoms (2 Cl atoms, for example) would cause more deshielding than only 1 Cl
atom.
 CHEMICAL SHIFT
 We define the relative position of absorption in the NMR spectrum the chemical
shift. It is a unitless number (actually a ratio, in which the units cancel), but we
assign ‘units’ of ppm or δ (Greek letter delta) units.
 For 1H, the usual scale of NMR spectra is 0 to 10 (or 12) ppm (or δ).
 The usual 13C scale goes from 0 to about 220 ppm.
 The zero point is defined as the position of absorption of a standard,
tetramethylsilane (TMS):
 This standard has only one type of C and only one type of H.
 The difference between the absorption position of a proton and the absorption
position of the protons of reference compound.
 Tetramethylsilane (TMS)
 TMS is added to the sample as internal standard.
 TMS protons are all identical, highly shielded providing a
single sharp peak always isolated from peaks of interest. The
TMS was assigned d = 0.00.
 Organic protons absorb downfield (to the left) of the TMS
signal.
 TMS is inert , highly soluble in organic liquids and easily
removed from samples by distillation.
Factors affecting chemical shift
1. Deshielding by electronegative elements Inductive effect by
electronegative groups
2. s-character (hybridization effect)
3. Magnetic Anisotropic effect (magnetic fields usually due to П-bonded
electrons in the molecule)
4. Hydrogen bonding.
1.DESHIELDING BY ELECTRONEGATIVE ELEMENTS
 ELECTRONEGATIVITY
DEPENDENCE OF CHEMICAL SHIFT

SUBSTITUTION EFFECTS ON
CHEMICAL SHIFT
 2. s-CHARACTER : (HYBRIDIZTION OF CARBON ATOM)
 As s-character of carbon atom increases, the electronic cloud is held more closer to the carbon and
provides less electron density for shielding of protons, and thus the chemical shift, δ, increases (shifted
downfield).
 According to the above reasoning, the following trend for the chemical shift is expected:

 This suggested that other factors than the sp character of carbon might affect the chemical shift in this
case. The above discrepancy can be explained by what is called Magnetic Anisotropic Effect.
3. MAGNETIC ANISOTROPIC FIELDS
 The presence of a nearby pi bond or pi system greatly affects the chemical shift.
Induced magnetic fields due to the П - electrons have greatest effect.
 In a magnetic field, the six П electrons in benzene circulate around the ring creating a
ring current.
 The magnetic field induced by these moving electrons reinforces the applied magnetic
field in the vicinity of the protons.
 The protons thus feel a stronger magnetic field and a higher frequency is needed for
resonance. Thus they are deshielded and absorb downfield.
 In a magnetic field, the loosely held П electrons of the double bond create a
magnetic field that reinforces the applied field in the vicinity of the protons.
 The protons now feel a stronger magnetic field, and require a higher frequency for
resonance. Thus the protons are deshielded and the absorption is downfield
 In a magnetic field, the pi electrons of a carbon-carbon triple bond are induced
to circulate, but in this case the induced magnetic field opposes the applied
magnetic field (B0 ).
 Thus, the proton feels a weaker magnetic field, so a lower frequency is needed
for resonance. The nucleus is shielded and the absorption is upfield.
 The outer 12 protons experience a
ring current induced reinforcing
field and absorb down field from
TMS, whereas the inner six
protons are subject to an opposing
field, absorbing upfield from
TMS
4. HYDROGEN BONDING
Information from 1H-NMR spectra:

1. Number of signals: How many different types of protons in the

molecule.
2. Position of signals (chemical shift): What types of protons.
3. Relative areas under signals (integration): How many protons
of each type.
4. Splitting pattern: How many neighboring protons.
1. Number of signals: How many different types of hydrogens in the
molecule
 The number of NMR signals equals the number of different types of protons in a
compound.
 Protons in different environments give different NMR signals.
 Equivalent protons give the same NMR signal.
 Magnetically equivalent protons resonate at the same applied field.
 Magnetically equivalent protons are also chemically equivalent. Hydrogens are
equivalent if
 They are truly equivalent by symmetry. -or-
 They are bonded to same atom and that carbon atom can rotate freely at room
temperature to interchange the positions of the hydrogens making the equivalent
to the spectrometer.
Equivalence by symmetry Equivalence by Rotation
• In comparing two H atoms on a ring or double bond, two protons are equivalent only if they are cis
(or trans) to the same groups.
2. Position of signals
Chemical Shift - 6 regions of NMR spectrum
1H NMR—Intensity of Signals
 The area under an NMR signal is proportional to the number of absorbing
protons.
 An NMR spectrometer automatically integrates the area under the peaks,
and prints out a stepped curve (integral) on the spectrum.
 The height of each step is proportional to the area under the peak, which
in turn is proportional to the number of absorbing protons.
 Modern NMR spectrometers automatically calculate and plot the value of
each integral in arbitrary units.
 The ratio of integrals to one another gives the ratio of absorbing protons in
a spectrum. Note that this gives a ratio, and not the absolute number, of
absorbing protons
Spin-Spin Coupling(Splitting)
 According to Dirac model-
 The electrons in the intervening bonds b/w the two nuclei transfer spin
information from one nucleus to another by means of interaction b/w nuclear and
electronic spin.
 Spin-spin coupling occurs due to un-pairing of bonded electrons.
 The interaction of the spin of neighbouring nuclei in a molecule causes splitting
of lines in the NMR spectrum & process is known as spin-spin coupling.
 Splitting causes due to Difference in environment of absorbing proton with
respect to neighbouring proton. Difference in spin state & induced magnetic field
 Peaks are often split into multiple peaks due to magnetic interactions between
nonequivalent protons on adjacent carbons, The process is called spin-spin
splitting
 The splitting is into one more peak than the number of H’s on the adjacent
carbon(s), This is the “n+1 rule”
 The relative intensities are in proportion of a binomial distribution given by
Pascal’s Triangle
 The set of peaks is a multiplet (2 = doublet, 3 = triplet, 4 = quartet, 5=pentet,
6=hextet, 7=heptet…..)
Rules for spin-spin splitting
Protons that are farther than two carbon
Equivalent protons do not split each other atoms apart do not split each other.
Vicinal coupling Geminal coupling
Magnitude of coupling constant J
 The magnitude of the coupling constant, J, can vary from 0 to about 20
Hz.
 This represents an energy gap (E = hn) due to the interaction of the nuclei
within the molecule.
 It does not depend on the strength of the external field.
 J is related to the dihedral angle between bonds. J largest for 0 (eclipsed)
or 180 (anti), smallest for 90, intermediate for gauche.
Relaxation Process
• When the nuclei are exposed to RF, in strong magnetic field, the nuclei moves to
higher energy states.
• Some times the absorption process = to the number of nuclei in two states and
cause absorption signal to decrease to zero (Saturation – saturated spin system –
where the number of nuclei in each energy state is equal).
• Mechanism should exist where by nuclei in an exited state can revert back to
ground state, otherwise the energy levels would equalise rapidly and no further
absorption should occur.
• Relaxation process is of two types:
a) Spin-lattice or longitudinal relaxation
b) Spin-spin relaxation
Spin-lattice or longitudinal relaxation Spin-spin relaxation

 Mainly in solids
 Transfer of energy from a nucleus in
an excited state to the molecular  Mutual exchange of spins between
lattice (molecular structural an excited nucleus and a neighbour,
environment in which the nucleus is without altering spin state of the
situated irrespective of state – solid, system.
liquid or gas).
 T2 = efficiency of spin spin
 Spin cooling.
relaxation.
 Nucleus returns to ground state and
the lattice gains energy.  When two neighboring nuclei of the
same kind have identical precession
 The efficiency of spin lattice
relaxation (T1) is expressed as the rates, but are in different magnetic
half life required for the system to quantum states, the magnetic fields
establish ah equilibrium state. of each can interact to cause an
 T1 – function of brownian motion in interchange of states.
liquids and lattice vibration in solids.
INSTRUMENTATION
 The continuous wave (CW) instrument
 The pulsed fourier transform (FT – NMR)instrument
The most important parts of an FT-NMR instrument are:
 The magnet,
 The RF generator,
 The sample chamber or probe, (which not only houses the sample but also the
RF transmission and detection coils).
In addition, the instrument requires:
 A pulse generator
 An RF receiver,
 Lots of electronics, and a computer for data processing
A Simplified 60 MHz NMR Spectrometer (continuous
wave Instrument)
 Older low field, classical (e.g., 60 MHz) NMR instruments used a fixed RF and
a permanent magnet or electromagnet with a set of Helmholtz coils in the pole
faces of the magnet.
 These coils could be adjusted to vary the applied magnet field slightly by
passing a current through them, causing each chemically different nucleus to
come into resonance sequentially. The sample holder was placed in the
magnetic field.
 Two RF coils surrounded the sample so as to be orthogonal to each other and to
the applied magnetic field.
 One coil applied a constant RF frequency;
 The second coil detected the RF emission from the excited nuclei as they
relaxed.
 These systems were simple and rugged, but limited in resolution and capability.
 There are no longer any manufacturers of CW NMR instrumentation.
FT NMR
 Main features of a basic NMR include:
– A radio transmitter coil that produces a short powerful pulse of radio
waves
– A powerful magnet that produces strong magnetic fields
– The sample is placed in a glass tube that spins so the test material is
subject to uniform magnetic field.
-Solid samples are dissolved in a solvent that will not give a signal (CDCl3 ,
CCl4 etc)
– A radio receiver coil that detects radio frequencies emitted as nuclei relax
to a lower energy level
– A computer that analyses and record the data.
 Superconducting Magnet (SCM): A Big Stainless Steel
Dewar
 use persistent superconducting magnets to generate the B0 field;
 At low temperatures (less than 6 K, typically) the resistance goes
to zero – that is the wire(eg.Nb alloy) is superconducting;
 To maintain the wire in its superconducting state the coil is
immersed in a bath of liquid helium (4 K, expensive);
 “Heat shield” kept at 77 K by contact with a bath of liquid
nitrogen (cheap) to reduces the amount of liquid helium boils off;
 Vacuum flask so as to further reduce the heat flow.
shim coils

 High resolution NMR requires linewidths of 1 Hz or less

 Magnetic field across the sample must be homogeneous so that the
corresponding variation in the Larmor frequency is small.
 Surround the sample with a set of shim coils, each of which produces a
tiny magnetic field with a particular spatial profile to canceling out the
small residual inhomogeneities in the main magnetic field.
 SAMPLE PROBE
 The sample chamber into which the sample holder is
placed is called the probe in an NMR spectrometer.
 The probe holds the sample fixed in the magnetic field,
contains an air turbine to spin the sample holder while the
spectrum is collected and houses the coil(s) for
transmitting and detecting NMR signals.
 The probe is the heart of the NMR system.
 The most essential component is the RF transmitting and
receiving coil, which is arranged to surround the sample
holder and is tuned to the precession frequency of the
nucleus to be measured.
 Modern NMR probes use a single wire coil to both excite
the sample and detect the signal.
 The coil transmits a strong RF pulse to the sample; the
pulse is stopped and the same coil picks up the FID signal
from the relaxing nuclei.
RF Generation and Detection
 The RF radiation is generated by using an RF crystal oscillator.
 The output of the oscillator is amplified, mixed, and filtered to produce
essentially monochromatic RF radiation for an NMR instrument.
 The RF radiation delivered to the sample is pulsed.
 The FID signal in the time domain must be converted to a frequency
domain spectrum by application of a Fourier transformation or other
mathematical transformation.
Signal Integrator and Computer
 All NMR spectrometers are equipped with a signal integrator to measure
the area of peaks.
 All NMR spectrometers require a computer to process the data and much
of the instrument is under computer control.
 Computer control of the room temperature shim currents that control the
field homogeneity, of sample spinning rates, autosamplers, pulse
sequences, and many other aspects of the instrument operation is a
necessity.
CW INSTRUMENT FT-NMR

 The chemical shifts in this spectrum  Here a powerful but short bust of
are calculated from frequency energy, called a pulse, that excites all
differences from TMS and it is a the magnetic nuclei in the molecule
frequency domain spectrum. simultaneously.
 Peak generated by this instrument  The source is turned on and off very
have ringing, a decreasing series of quickly, generating a pulse.
oscillations that occurs after the  When the pulse is discontinued the
instrument has scanned through the nuclei relaxes by emitting EMR.
peak.  Since the molecule contains
 As a result they are still emitting an different nuclei, many different RF
oscillating, rapidly decaying signal, are emitted simultaneously. This is
recorded as ringing. FID.
Advantages of FT method over CW method
 More sensitive and can measure weaker signal
 5-10min required to record CW spectrum while FID can be performed in
few seconds.
 With a computer and fast measurement, possible to repeat and average the
measurement of the FID signal.
 Advantageous when the sample is small.
APPLICATIONS
STRUCTURE ELUCIDATION
 • Based on the four points discussed in the interpretation of the spectra.
INVESTIGATION OF DYNAMIC PROPERTIES OF MOLECULES
 Conformational isomerism
 Restricted rotation
 Molecular asymmetry
 Hydrogen bonding
 Keto enol tautomerism
 Determination of optical purity
 Molecular interactions
 Micelle formation
 Drug macromolecule interactions
 Quantitative analysis
THANK YOU