State of measurements for the various techniques

electronic UV-Vis

vibrational infrared

rotational microwave

nuclear spin radiofrequency

NMR is concerned with change in the direction of

spin orientation as the result of the absorption of
radiofrequency radiation.
 Nuclear Magnetic Resonance (NMR)
(MOST Routinely used Analytical Techniques)

Introduction:
Nuclear Magnetic Resonance (NMR) measures the absorption of
electromagnetic radiation in the radio-frequency region (~4-900 MHz)
- nuclei (instead of outer electrons) are involved in
absorption process
- sample needs to be placed in magnetic field to cause
different energy states

NMR- first experimentally observed (1946) by Bloch and Purcell (got

Nobel Prize in 1952) -quickly became commercially available and widely
used.

NMR can routinely and rapidly elucidate : Composition, Structure,

Dynamics and Function of the Chemical Entities (small organic
molecules to large molecular weight polymers and proteins).
 Introduction
• It is used to study a wide variety of nuclei:
1
H, 13C,15N,19F,31P
• A nucleus with an odd atomic number or an
odd mass number has a nuclear spin.
• The spinning charged nucleus generates a
magnetic field.
 NMR Spectroscopy
NMR study specific atomic nuclei
NMR applying exclusively to study magnetic properties of nuclei
with spin quantum s # > 0. The spin quantum number is
associated with the mass number and atomic number of the
nuclei as given in the following table.

The nucleus of H, the proton has s = ½ and is magnetic where as

14
C, and 16O have s = 0 and therefore are non-magnetic
 The NMR Phenomenon
The spinning Nucleus:

Spinning charged particles such as nucleus of the

hydrogen atom behave as a tiny spinning bar
magnet and generate their magnetic field.

Effect of external magnetic field:

On application of strong external magnetic field, the

spinning charge (say proton) is forced to precess
(wobbling motion). The precessional motion of the
proton either align parallel (having low energy state)
or antiparallel (having high energy state).
See the following figure
 Two Energy States
A photon with the right
amount of energy can be
absorbed and cause the
spinning proton to flip from
the parallel (low energy
state) to antiparallel (high
energy state) .

Energy difference between

the two state is
proportional to the
magnetic field strength
 The NMR Phenomenon cont’d
Energy transitions:

Irradiating the precessing nucleus with a beam of

radiofrequency energy of the correct frequency, the low energy
nucleus may absorb this energy and move to a higher energy
level. In aligned orientation, the proton will absorb energy and
will pass to the opposed orientation. Subsequently, it will lose
this extra energy and come back in to the aligned state. The
precessing proton will only absorb energy from the
radiofrequency source if the precessing frequency is the same
as that of the proton i.e. in resonance hence the term nuclear
magnetic resonance. The absorption of the energy in the form
of NMR Graph is recorded.
 Nuclear Spin
NMR spectroscopy employs an magnetic energy absorption
process, which orients spinning nuclei in a strong external
magnetic field.

𝟏 𝟏𝟒
𝐇 𝐍
NMR spectroscopy observes isotopes having odd mass numbers
and/or odd atomic numbers.*
 Nuclear Spin of protons and its separation to two groups
On application of a strong external magnetic field is applied, the
protons with spin aligned to the external field and non-aligned
separate out as compared to the absence of this field when it is
random as: 𝜷
+¿ ¿ +¿ ¿
P P
+¿ ¿
P
+¿ ¿
P
alignment opposes the applied magnetic
field and is higher in energy than
∆𝐄
+¿ ¿ alignment.

+¿ ¿ P
P
+¿ ¿ +¿ ¿
P P
𝐇𝟎 𝜶
No applied magnetic field Influence of applied magnetic field

Manipulation of nuclei is through the application of an applied magnetic field, .

In the example, the nucleus, such as 1H, has a nuclear spin of ½.
 Magnetic Shielding
• If all protons absorbed the same
amount of energy in a given magnetic
field, not much information could be
obtained.
• But protons are surrounded by
electrons that shield them from the
external field.
• Circulating electrons create an induced
magnetic field that opposes the external
magnetic field.
 Shielded Protons
Magnetic field strength must be increased
for a shielded proton to flip at the same
frequency.
 Protons in a Molecule
Depending on their chemical
environment, protons in a molecule are
shielded by different amounts.
 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.
The NMR Graph
Tolluene have two types of hydrogens i.e. hydrogens
(3) in methyl group of toluene and hydrogens (5) as a
part of the benzene ring thus giving two signals with
intensity ration 5:3.
 CH3
H3C Si CH3 Tetramethylsilane
CH3

• TMS is added to the sample.

• Since silicon is less electronegative
than carbon, TMS protons are highly
shielded. Signal defined as zero.
• Organic protons absorb downfield (to
the left) of the TMS signal.
 Chemical Shifting
Chemical shift is the position of signals in NMR spectra due to
various chemical influences (environments) within the
molecule.
Chemical Environments
The magnitude of
Chemical environment is the a chemical shift is
molecular arrangement existing determined by the
around a given proton. chemical
environment of a
Variation in chemical H H H given proton.
environment of atoms
within the molecules
is responsible for Br
C C C Br

variation in its
response to the H H H
applied field.
 Chemical Shift
The magnitude of variation in chemical environment of
atoms within the molecules which is responsible for
variation in its response to the applied field is measured as
chemical shift.
• Chemical shift is measured in parts per million (ppm).
• 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.
NMR spectrum give information as:
1. number of signals
2. their intensity (as measured by area under peak)
3. splitting pattern (multiplicity)
 Delta Scale

It is clear that the variation in strength of the external

field does not changes the values of the chemical shift
Location of Signals (chemical shift)
• More electronegative
atoms deshield more and
give larger shift values.
• Effect decreases with
distance.
• Additional electronegative
atoms cause increase in
chemical shift.
Typical chemical shift values

Chapter 13
The chemical shift of the signals of the spinning nuclei in
the NMR spectrum depends upon various factors as:
 Chemical Shifting

These specialized environments are a creation of

the influences of several factors:
Understanding the
chemical environment is
important in NMR
spectroscopy because the
hybridizatio chemical environment
influences the positioning
n
Electronegativit of the proton signals in
y the 1H NMR spectrum.
Hydrogen-
bonding

Induced magnetic
fields
These factors affect chemical
environments by either interacting
Applied magnetic with the electron density surrounding
field the proton or the magnitude of the
Electron applied magnetic field felt by the
Density proton.
 Chemical Shifting-Induced magnetic affect
When nuclei are excited by an external applied field, the electrons
surrounding the nuclei can produce individual magnetic fields with anti
parallel magnetic affect also called shielding affect that oppose the applied
field.
When a circulating electron produces a magnetic field aligned with the
external field, it enhances the external field’s affect on the proton. This is
known as deshielding.
Shielded protons will appear upfield and deshielded protons will
appear downfield.
Deshielded Shielded

−
𝑒 𝑒
−

5.0 4.0 3.0 2.0 1.0

Down field Up field

 Chemical Shifting-Electron density

Increase in density of the electron cloud will cause a proton to appear upfield
in the NMR spectrum.
Likewise, decrease in electron cloud will cause protons to appear
downfield on the NMR spectrum.
Apart from variation in electron cloud, the chemical shift of the proton in the
spectrum is also dependent on the magnitude of distortion in the electron
cloud.

H
H
H

5.0 4.0 3.0 2.0 1.0

Down field Up field

 Chemical Shifting - Electronegativity

The magnitude of
influence bonded atoms
H have on a proton will be
dependent on the extent
O of the electronegativity of
H H
the atom, or groups of
C C atoms, and the atom(s)
O bonding position is in
C C
respect to the proton.
H H

H C N O F
Increasing
 Chemical Shifting-Functional Groups

Amide Functional groups are

Understanding the impact
groups of atoms (or
of electronegativity on
individual atoms) that
electron density
N replace hydrogen by
necessitates the need to
covalently bonding to the
know functional groups.
carbon framework.

Alcohol

H
O
O
Carboxylic
Acid

The kinds of atoms which

The more
replace hydrogen will
F electronegative atoms
dictate the extent of
will produce bigger
electron cloud distortion.
distortions.
Halide
 Chemical Shifting-Electronic cloud

The distortion of the electron cloud

surrounding a proton creates unique
characteristics and behaviors of the
molecule insomuch that by their presence
they form classes of organic compounds.

Functional Groups appearing on NMR

spectra due to chemical shifting.
 Chemical Shifting- Hydrogen-Bonding

An extreme interaction of electronegativity is hydrogen-bonding.

+¿¿
𝛿
− 𝛿 Due to the strong electronegative behavior of
hydrogen-bonding, the distortion of the
electron density of neighboring protons is
greater than atoms or functional groups who
do not posses the ability to hydrogen-bond.

+¿¿ The ability for neighboring atoms

𝛿 to undergo hydrogen-bonding
means an adjacent proton will
have less electron density around
it.

−
𝛿
 Multiplicity

Signals often contain two or three or

more peaks due to proton-proton
splitting. Therefore signals may appear
as a group of peaks.

Multiple peaks in a signal occur due 𝐂𝐇𝟑

to the interaction between adjacent
or neighboring protons.

The influences between protons arises

because neighboring protons interact with
each other’s magnetic fields.

OH 𝐂𝐇𝟐
This phenomenon is
called

Multiplicity
 Chemical Shifting

The closer electronegative atom(s) are to a proton, the greater

the downfield shift of the proton.

O
H H
A proton located more than three
sigma bonds is too far away to
C C interact and therefore will not
H cause any downfield shift.
3 1 O
C C

H 4 H H 2 H

Therefore, protons on carbon 2 will feel a larger decrease in their electron density than
protons on carbon 3. Carbon 4 protons will not be affected.
 Electronegative substituents decrease
the shielding of methyl groups

CH3F d 4.3 ppm

CH3OCH3 d 3.2 ppm
CH3N(CH3)2 d 2.2 ppm
CH3CH3 d 0.9 ppm
CH3Si(CH3)3 d 0.0 ppm CH3OCH3 CH3CH3

CH3F CH3N(CH3)2 CH3Si(CH3)3

 Effect is cumulative

CHCl3 d 7.3 ppm

CH2Cl2 d 5.3 ppm
CH3Cl d 3.1 ppm

CHCl3 CH3Cl

CH2Cl2
 Number of Signals
Equivalent hydrogens have the same
chemical shift.

Chapter 13 37
 Intensity of Signals
• The area under each peak is
proportional to the number of protons.
• Shown by integral trace.
 How Many Hydrogens?
When the molecular formula is known,
each integral rise can be assigned to a
particular number of hydrogens.
 4.1 Applications of NMR Spectroscopy

Drug
Solution Structure
Screening & Design

Molecular Chemical Analysis

Dynamics

Applications of
Intermolecular
NMR Interactions
Spectroscopy
Material Science

Protein
Metabolite Analysis Hydration

Protein Folding
Ionization State
Native Membrane
Protein