NMR

SPECTROSCOPY
Afsath.
B M Pharm 1st
year
Pharmacognosy and Phytochemistry
• Spectroscpy is the study of interaction of EMR with matter, which may result in
absorption, transmission, emission,reflection , rotation of EMR
• Nuclear magnetic resonance spectroscopy, most commonly known as NMR spectroscopy.
Nuclear Magnetic Resonance (NMR) is a spectroscopy technique which is based on the
absorption of electromagnetic radiation in the radio frequency region 4 to 900 MHz by nuclei of
the atoms.
• Proton Nuclear magnetic resonance spectroscopy is one of the most powerful tools for elucidating
the number of hydrogen or proton in the compound.
• Spectroscopy determines the physical and chemical properties of atoms or the molecules in which
they are contained and provide detailed information about the structure, dynamics, reaction state, and
chemical environment of molecules.
• It is used to study a wide variety of nuclei:
• 1H
• 15N
• 19F
•• 31
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13C
P
 Theory of
NMR
Spin quantum number (I) is related to the atomic and mass number of the
nucleus I Z A Eg;

Half integer Odd Odd 1H (1/2)

Half integer Odd Even 13C (1/2)

Integer Even Odd 2H(1)

Zero Even Even 12C (0)

Elements with either odd mass or odd atomic number have the property of nuclear
“spin”
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• If an external magnetic field is applied,the number of possible orientations calculated by
(2I+1).
Eg.:-Hydrogen has spin quantum number I=1/2 and possible
orientation is (2*1/2+1=2) two , ie,+1/2 and -1/2.

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 PRINCIPLE OF
•The principle is based on the-
NMR
spinning of nucleus and
generating a magnetic field.
•Without external magnetic(Bo)
– field nuclear spin are random
in direction.
•With Bo ,nuclei align themselves
either with or against field of
external
magnetic field
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Eg, Liq.N2 Vs Liq.
O2

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• If an external magnetic field is applied, an energy transfer (ΔE) is possible between ground
state to excited state.
• When the spin returns to its ground state level, the absorbed radiofrequency energy is emitted
at the same frequency level.
• The emitted radiofrequency signal that give the NMR spectrum of the concerned nucleus.and
is
directly proportional to the strength of the applied field

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 NMR
SPECTRUM
NMR spectrum is a plot of intensity of NMR signals VS magnetic field (frequency) in
reference to TMS

Reasons for taking TMS as reference

Standard
1. Chemically inert, magnetically isotopic, volatile & soluble in most organic
solvents.
2. TMS gives an intense signal.
3. TMS can be easily removed.
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4. Electro negativity is low.

 Chemical
• Is the resonance frequency of a nucleusshift
relative to a standard in a magnetic field.
• TMS is the most common reference compound in NMR, it is set at 𝛿 = 0𝑝𝑝𝑚
𝛿=ν𝑠𝑎𝑚𝑝𝑙𝑒 − 𝜈𝑟𝑒𝑓
𝜈𝑟𝑒𝑓
Shielding of protons:- High electron density around a nucleus shields the nucleus
from the external magnetic field and the signals are upfield in the NMR spectrum
Deshielding of protons:- Lower electron density around a nucleus deshields the
nucleus from the external magnetic field and the signals are downfield in the NMR
spectrum

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 Factors affecting chemical
• Electronegative groups/ Inductive
shift effect
• Magnetic anisotropy of π-systems / diamagnetic effect of pi bond
• Hydrogen bonding
• Vander Waal's deshielding
• Effect of temperature &Effect of solvent
Electronegative groups:- Electronegative groups attached to the C-H system
decrease the electron density around the protons, and there is less shielding
(i.e.deshielding) and chemical shift increases
Compound Chemical shift
CH3I 2.16

CH3Br 2.65
CH3Cl 3.10 10

CH3F 4.26
Magnetic anisotropy of π-systems-( space
effect)
 So magnetic anisotropy means that there is a "non-uniform magnetic field".

 Electrons in π systems (e.g. aromatics, alkenes, alkynes, carbonyls etc.) interact with the applied
field which induces a magnetic field that causes the anisotropy.

 It causes both shielding and de-shielding of

protons. Example:-Benzene
Vander Waal's deshielding
 The electron cloud of a bulkier group will tend
to repel the electron cloud surrounding the
proton.
 such a proton will be deshielded & will resonate at slightly higher value of δ than
Effect of temperature
expected in the absence of this effect. 11

Resonance position of most signals is little affected by

Effect of solvent
Chemical shift change when the solvent changed from CCl4 to CDCl3 is 0.1 ppm. But
change to more polar solvents like methanol, the change is 0.3 ppm. Solvents used in
CCl4, CS2, CDCl3, C6D6, D2O
NMR
Hydrogen bonding
 Protons that are involved in hydrogen bonding are typically change the chemical shift
values.
 The more hydrogen bonding, the more proton is deshielded and chemical shift value is
higher.

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 Proton
The most common for of NMR is based
NMRon the hydrogen-1 (1H), nucleus or proton. It
can give
information about the structure of any molecule containing hydrogen atoms.
Eg, ethanol
3 types of ;-CH2,CH3,OH

Interpretation of 1HNMR
spectra:- - Indicates how many "different kinds" of protons are
Number of present.
signals Position - Indicates something about (chemicalshift)
Relative
of signalsintensity of signals magnetic (electronic) environment of protons
Splitting of signals (spin spin - Proportional to number of protons present signals
coupling) - Indicates the number of nearby nuclei
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n+1
rule
• The multiplicity of signal is calculated by using n+1 rule.
• This is one of the rule to predict the splitting of proton signals. This is considered
by the nearby hydrogen nuclei.
Therefore, n= Number of protons in nearby nuclei
 Zero H atom as neighbour n+1=0+1=1(singlet)
 One H atom as neighbour n+1=1+1 = 2(doublet)
 Two H atom as neighbour n+1=2+1 =3(triplet)
Spin-spin coupling (splitting)
The interaction between the spins of neighbouring nuclei in a molecule may cause
the splitting of NMR spectrum. The splitting pattern is related to the number of
equivalent
atom H-nearby nuclei. Eg., Ethyl
at the
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acetate
Spin-spin coupling (splitting)
cont…

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 Rules for spin-spin
• coupling
Chemically equivalent protons do not show spin-spin coupling &Only nonequivalent protons
couple. X
X Hb Hc Hd H Hb couples with Hc
Ha C C C C Hb & Ha donot couple because they are equivalent
HH Hc & Hd donot couple because they are
H equivalent
H
• Protons separated byHfour or more bonds will not
couple.
• Protons on adjacent carbons normally will
couple.

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ORIGINS OF SIGNAL
SPLITTING
The origins of signal splitting patterns in
which, each arrow represents an Hb
nuclear spin orientation.

COUPLING CONSTANT (J-Hz)

• Measurement of splitting effect is based
on the distance between the peak in a
given multiplet.
• Useful in 1H NMR of complex structure 17
 INSTRUMENTATION
1. Sample tube/sample
holder
2. Permanent magnet
3. Magnet coil
4. Sweep generator
5. Radio frequency
transmitter
6. Radio frequency
reviever
7. Read out system

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• Sample tube / sample holder
It should be chemically inert, durable & transparent to NMR radiation.
Generally about 8.5 cm long & approximately 0.3 cm in diameter is
employed.
• Sample probe
It’s the device that hold sample tube in position & is provided with an air driven
turbine for rotating the sample tube almost 100 revolutions per min.
• Permanent Magnet
It provide homogenous magnetic field at 60-100MHz
• Magnetic coil
It induce magnetic field when current flow through them
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• Sweep generator
• Radio frequency transmitter
transmitter is fed on to a pair of coils mounted on right angles to the path of
field. 60 MHz capacity is normally used.
• Radio frequency receiver
detect radio frequencies emitted as nuclei relax at lower energy level
• Signal detector & recording system
The electrical signal generated is amplified by
means of amplifier & then recorded.

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Application
………
• 1H NMR used for structural elucidation of organic and inorganic
solids

• determines the physical and chemical properties of atoms

• Application in medicine

• Anatomical imaging

• Measuring physiological function

• Flow measurement and angiography

• Tissue perfusion studies

• Tumours 21

• MRI
Application in
medicine… BRAIN
• Distinguishing grey matter & white matter
• Imaging posterior fossae , brain stem, spinal cord
• Detect demyelinating lesions, tumour, haemorrhage, infarctions
ABDOMEN
• Metabolic liver disease
• Focal areas of inflammation in chronic active hepatisis
KIDNEY
• Distinguishing renal corta & medulla
• To evaluate transplanted kidney
PELVIS 22

• Differentiate between benign prostatic hyperplasia &prostatic

 REFERENCE
• Organic spectroscopy, Willaim Kemp
• Instrumental method of chemical analysis
• Instrumental method of chemical analysis. Wilard
• https://www.slideshare.net/solairajananant/nmr-spectroscopy-13887430
• http://sydney.edu.au/science/chemistry/facilities/nmr/nmr-applications.shtml
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