NMR Spectroscopy

Dedicated to
Team D
Dr. Md. Elius Hossain
Sadnan Chowdhury
Assistant Professor
Aruna Ayfar
Department of Chemistry
Md. Yeasin Arafat Tarek
Bangladesh University of Engineering &
Raisul Awal Mahmood
Technology (BUET)
Rezwan Tarek
 Table of content
1. Electromagnetic radiation
2. History
3. What is Nuclear Magnetic Resonance (NMR)
4. Criteria for NMR technique
5. Instrumentation: Basic Operation of NMR Spectrometer
6. Solvents in NMR technique
7. Shielding & Deshielding
8. Chemical Shift
Electromagnetic Radiation (EMR)
NMR spectroscopy is studied
with the help of this region
History of Nuclear Magnetic Resonance (NMR)

Swiss-American Physicist Felix Bloch (left)

and American Physicist Edward Mills
Purcell (right) in 1950 who first
successfully detected an NMR signal and
Photo of Bloch's probe with the cover plate removed (left) and electromagnet got the Nobel Prize in 1952.
with the probe containing a sample tube about to be inserted into the magnet
gap (right). (From Bloch's Nobel Prize address, entitled "The Principle of
Nuclear Induction," reprinted with permission from Science, 1953, 118, 425.)

Ref. Analytical Chemistry, Vol. 65, No. 6, March 15, 1993

What is Nuclear Magnetic Resonance (NMR)
NMR is a physical phenomenon to investigate
molecular properties of matter by irradiating
atomic nuclei in a magnetic field with the source
of long wavelength radio wave energy. So its
corresponding frequency and energy are both
low. When these low-energy radio waves interact
with a molecule, they can change the nuclear
spins of some elements, including 1H and 13C.
In a magnetic field, there are two different
energy states for a proton:
• A lower energy state with the nucleus aligned
in the same direction as B0
• A higher energy state with the nucleus aligned
opposed to B0
The stronger the magnetic field, the larger the energy
difference between the two nuclear spin states, and
the higher the m needed for resonance.
A nucleus is in resonance when it
absorbs RF radiation and “spin
flips” to a higher energy state.
Nuclear Magnetic Resonance (NMR) Technique
In the absence of an external magnetic field,
the spins of magnetic nuclei are oriented
randomly. In the figure A, Nuclear spins are
oriented randomly in the absence of an
external magnetic field
oriented randomly
Figure A
When a sample containing these nuclei is
placed between the poles of a strong magnet,
the nuclei adopt specific orientations shown in
figure B, much as a compass needle orients in
the earth’s magnetic field.
Applied magnetic field, 𝑩𝑩𝟎𝟎
specific orientations
Applied magnetic field, 𝑩𝑩𝟎𝟎
Figure A
Nuclear Magnetic Resonance (NMR) Technique
A spinning 1H or 13C nucleus can orient so that its
own tiny magnetic field is aligned either with N
(parallel to) or against (antiparallel to) the external
field. The two orientations don’t have the same
energy, however, and aren’t equally likely. The
parallel orientation is slightly lower in energy by an Radio wave
amount that depends on the strength of the external
field, making this spin state very slightly favored over
the antiparallel orientation.
External
Higher energy state
magnetic
field

Absorb energy

Lower energy state S

The energy difference ∆𝑬𝑬 between nuclear spin states depends on the strength of Absorption of energy with frequency converts a nucleus from a
the applied magnetic field. lower spin state to a higher spin state.
Criteria for Nuclear spin in NMR Spectroscopy
 All nuclei with an odd number of protons (1H,
2H, 14N, 19F, 31P) show magnetic properties. Mass Atomic Spin Examples
Number Number Number
 All nuclei with an odd number of neutrons (13C) 𝟏𝟏 1H, 13C, 15N,
show magnetic properties. Odd 𝟐𝟐 19F, 31P

 Only nuclei with even numbers of both protons Odd Or, 𝟑𝟑 11B, 35Cl, 37Cl,

and neutrons (12C, 16O, 32S) do not give rise to Even 𝟐𝟐 79Br, 81Br

𝟓𝟓 127I, 17O
magnetic phenomena.
𝟐𝟐
 Practically, field strengths in the range of 4.7 to 12C, 16O, 32S,
7.0 tesla (T) are more common. At a magnetic Even Even 𝟎𝟎 34S

field strength of 4.7 T, so-called radiofrequency

energy in the 200 MHz range (1 MHz = 106 Hz) 14N, 2H, (or
𝟎𝟎 D)
brings a 1H nucleus into resonance, and 50 MHz
Even Odd
brings a 13C nucleus into resonance. At the
highest field strength of 1000 MHz energy is 3 10B

required for 1H spectroscopy.

Instrumentation: Basic Operation of NMR Spectrometer
The operation of a basic NMR spectrometer is
illustrated in the figure.
 A sample is dissolved in a suitable solvent
(usually deuteriochloroform, CDCl3, which has
no hydrogens) and placed in a thin glass tube
between the poles of a magnet.
 The strong magnetic field causes the 1H and 13C
nuclei in the molecule to align in one of the two
possible orientations and the sample is irradiated
with radiofrequency energy. If the frequency of
the radiofrequency irradiation is held constant Fig. Schematic operation of a basic NMR spectrometer. A
thin glass tube containing the sample solution is placed
and the strength of the applied magnetic field is between the poles of a strong magnet and irradiated with
varied, each nucleus comes into resonance at a radiofrequency energy.
slightly different field strength. Note: All protons do not absorb at the same
 A sensitive detector monitors the absorption, frequency. Protons in different environments
and its electronic signal is then amplified and absorb at slightly different frequencies, and so
displayed as a peak. they are distinguishable by NMR.
A brief view of NMR spectrum
1H NMR spectrum analysis

Radiation Magnetic field Sample Detector Recorder

Radio- 60 Hz (Solid, Liquid)
frequency 4-7 tesla (T)

NMR spectrum

Number of Signals Signal Area Chemical Shift Spin-spin coupling

Set of Total no. of Identification No. of proton in

equivalence proton in a of different neighboring
protons structure groups carbon (n+1 rule)
Solvents in NMR technique
In NMR spectroscopy, usually proton-less solvents
are used so that no peak is observed for that
particular solvent. For example – CCl4, CS2, CDCl3,
D2O, C6D6, (CD)3CO etc.
Characteristics:
 Solubility: Higher solubility of sample in the
chosen solvent to contribute the higher
sensitivity.
 Purity: Degree of pure solvent is a major
selection criteria to avoid the interference of
solvent peaks with sample peaks
 Viscosity: Lower viscosity to get better spectral
resolution.
 Moisture free: Moisture contents should be free
to give better peak signals.
 Deuterated solvents: Deuterated solvents should
be used to minimize the interference of solvents
and sample peaks.
Tetramethyl silane (TMS) is used as reference
solvent in the sample mixture to understand
the chemical shift of the particular sample.
Characteristics:
 Highly soluble in most organic solvents.
 Chemically inert to the sample
 Very volatile, so it can be volatized from the
sample.
 Contains 12 equivalent 1H and so shows a
single peak even in low concentration.
 TMS signal is set to 0.
 Insoluble in water, so the samples which are
insoluble in water, can not be used with
TMS. CH 3
H3C Si CH3 Tetramethyl silane (TMS)
CH3
Shielding & Deshielding
An electron around the nucleus creates a small
magnetic field that opposes the applied
magnetic field, B0. Thus the electron is said to A proton surrounded by electron
density
shield the nucleus from B0. The magnetic field
An isolated proton
generated by the circulating electron decreases
the external magnetic field that the proton
“feels.” Because the proton experiences a lower
magnetic field strength, so, it needs lower
frequency to achieve resonance.

The nucleus “feels” Bo only. Magnetic field induced by the

Shielded protons absorb radiation at higher electron (opposite to Bo). The
induced field decreases the
fields (frequency) while the deshielded protons strength of the magnetic field
will absorb at lower fields (frequency). “felt” by the nucleus. So, The
nucleus is shielded
Chemical Shift
The chemical shift of a nucleus is the difference
between the resonance frequency of the nucleus
and a standard, relative to the standard. In other
words, Chemical shift is a measure of the degree
to which a nucleus in a molecule is shielded or
deshielded.
The shielding of protons in different
environment are different, so they have different
chemical shift
Characteristics:
• Measured in parts per million (ppm).
• Ratio of shift downfield from TMS (Hz) to
total spectrometer frequency (MHz).
• Has the same value regardless of the
machines (same value for 60, 100, or 300
MHz machine).
• Called the delta (𝛿𝛿) scale ranged from 0 to 13.
Chemical Shift
𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇 𝒐𝒐𝒐𝒐 𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔 − 𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇 𝒐𝒐𝒐𝒐 𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔
𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪𝑪 𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔 = × 𝟏𝟏𝟏𝟏𝟔𝟔 𝒑𝒑𝒑𝒑𝒑𝒑
𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔𝒔 𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇𝒇
Factors that affect Chemical Shift, 𝜹𝜹
Electronegativity – δ- δ+
As chlorine is more electronegative atom, so
chlorine “deshields” the proton, that is, it takes
valence electron density away from carbon, which Cl C H
in turn takes more density from hydrogen δ- δ+
Electronegative atom
deshielding the proton.

NMR Chart
“Deshielded“ “Highly shielded”
protons appear protons appear
at downfield at upfield
(higher δ) (lower δ)

deshielding moves proton

resonance to lower field and higher δ
Factors that affect the Chemical Shift, 𝜹𝜹
Electronegativity (Dependence of the Chemical Shift of CH3X on the X atom) –
Compound CH3X CH3F CH3OH CH3Cl CH3Br CH3I CH4 (CH3)4Si
Element X F O Cl Br I H Si
Electronegativity of X 4.0 3.5 3.1 2.8 2.5 2.1 1.8
Chemical shift, 𝜹𝜹 4.26 3.40 3.05 2.68 2.16 0.23 0
TMS
Most deshielded Deshielding increases with the electronegativity of atom X

Substitution effects on Chemical Shift –

CHCl3 CH2Cl2 CH3Cl The effect increases with greater numbers of

Most deshielded electronegative atoms
7.27 5.30 3.05 ppm
The effect decreases with increasing
−CH −Br −CH −CH −Br − CH −CH −CH −Br distance from the electronegative atom.
Most 2 2 2 2 2 2
The effect completely vanished at the
deshielded 3.30 1.69 1.25 ppm fourth bond from the electronegative
atom.
Factors that affect Chemical Shift, 𝜹𝜹
Hybridization of carbon atom & Anisotropy – But in fact, we get the following order,
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 C H > C H > C H
protons, and thus the chemical shift, δ increases
sp2 sp sp3
(shifted downfield).
actually observed
According to the above reason, the following
at δ = 2-3
trend for the chemical shift is expected:

C H > C H > C H This suggested that other factor might affect

sp sp2 sp3 the chemical shift in this case. The above
discrepancy can be explained that leads to
s-character increases
Magnetic Anisotropic Effect.
(down field - higher δ)
Expected to be
observed at δ > 7
Factors that affect Chemical Shift, 𝜹𝜹
Isotropy –
When the properties of a molecule are the same in
all direction frames or points, then it is called
isotropy.
Anisotropy –
When the properties of a molecule are the
different in all direction frames or points, then it
is called anisotropy. Anisotropy is direction Isotropy Anisotropy
dependent. Shielded Induced
region magnetic
field

In case of molecule, valence electrons shield the

nucleus from the full effect of the applied field. Deshielded Deshielded
region region
The applied field induces circulation of the
valence electrons and opposes the applied field.
Shielded
region
 Induced
Factors that affect Chemical Shift, 𝜹𝜹 magnetic field
Shielded
region
Anisotropy effect –
Due to have presence of π- electrons, induced magnetic Deshielded
region
fields have greatest effect on the conjugate system. The
presence of a nearby π- bonds or π- system greatly affects
the chemical shift.
 Highly deshielded means chemical shift is in downfield
 Highly shielded means chemical shift is in upfield
Aromatic protons, δ = 7-8 ppm
Induced Induced
magnetic Shielded region magnetic field Induced
Shielded Shielded
field magnetic
region region
field
Deshielded
region

Deshielded
region

Deshielded
region

Acetylene protons δ̃ ≈ 2.5 ppm Vinyl (Olefinic) protons, δ = 5-6 ppm Aldehyde proton, δ= 9-10 ppm
Factors that affect Chemical Shift, 𝜹𝜹
R
Hydrogen bonding –
The chemical shift depends on how much O H H
hydrogen bonding is taking place (observed in
high concentrated solutions). Hydrogen bonding
O H O R
lengthens the O-H bond and reduces the valence
electron density around the proton R
 H-bonding deshields the protons and shifts the
Alcohols vary in chemical shift from 0.5 ppm
NMR spectrum in downfield. (free OH) to about 5.0 ppm (lots of H
δ- δ+ δ- bonding).
O H O O O
-I
R C C R H 3C NH Intramolecular H-bond:
In methyl salicylate,
which has strong internal
O H O O hydrogen bonding, the
δ- δ+ δ- NMR absorption for O-H
is at about 14 ppm,
Intermolecular H-bond: Due to formation of hydrogen bonding, the (highly downfield)
protons attached to carboxylic acids forming dimer are the least
shielded protons and have a chemical shift of 10-12 ppm.
Factors that affect Chemical Shift, 𝜹𝜹
Steric hindrance
van der Waals deshielding–
In the overcrowded molecules, it is possible that
some proton may be occupying steric hindered
H
position. R
Electron crowd of a bulky group will tend to repel
the electron cloud (in over crowded molecules or
hindering groups) surrounding the proton.

Such proton will be deshielded and will resonate

in lower field with high chemical shift. O

 When 𝑹𝑹 = 𝑯𝑯, no hindrance, so the  When 𝑹𝑹 = 𝑪𝑪𝑯𝑯𝟑𝟑 , hindrance governs, so

peak will appear in upper field with the peak will appear in down field with
low chemical shift. high chemical shift.
NMR Correlation Chart
Chemical shift gives the electronic environment of protons (Shielding and Deshielding)

Disappears upon shaking with D2O

Aromatic -OH -NH

CHCl3 H 0-5
3-5

Deshielded 10-12 7-8 1.5 - 3

TMS
Shielded
9-10 4.5 - 7 0–2
Downfield Upfield
High frequency 12 11 10 9 8 7 6 5 4 3 2 1 0 𝜹𝜹 (ppm) Low frequency
Large δ Small δ
H
CH2F CH2Ar C-CH-C
RCOOH RCHO C=C CH2Cl CH2NR2
CH2S C
Acid Aldehyde CH2Br
Olefinic C-CH2-C
CH2I C C-H
sp2 C-CH3
CH2O C=C-CH2
CH2NO2 CH2-C- sp3
sp3 sp sp3 O
Approximate Chemical shift ranges (ppm) for selected types of protons
R-CH3 0.7 - 1.3 R-N-C-H 2.2 - 2.9 R-C=C-H
R-CH2-R 1.2 - 1.4 R-S-C-H 2.0 - 3.0 4.5 - 6.5
R3CH 1.4 - 1.7
I-C-H 2.0 - 4.0
R-C=C-C-H 1.6 - 2.6 H
Br-C-H 2.7 - 4.1
O
6.5 - 8.0
R-C-C-H 2.1 - 2.4 Cl-C-H 3.1 - 4.1
O
O RO-C-H 3.2 - 3.8 R-C-N-H
RO-C-C-H 2.1 - 2.5 5.0 - 9.0
HO-C-H 3.2 - 3.8 O
O
O
HO-C-C-H 2.1 - 2.5 R-C-H
R-C-O-C-H 3.5 - 4.8 9.0 - 10.0
N C-C-H 2.1 - 3.0 O
O2N-C-H 4.1 - 4.3
R-C C-C-H 2.1 - 3.0 R-C-O-H
F-C-H 4.2 - 4.8 11.0 - 12.0
C-H 2.3 - 2.7 R-N-H 0.5 - 4.0 Ar-N-H 3.0 - 5.0 R-S-H
R-C C-H 1.7 - 2.7 R-O-H 0.5 - 5.0 Ar-O-H 4.0 - 7.0 1.0 - 4.0
Information from NMR spectrum

NMR spectrum

Number of Signals Signal Area Chemical Shift Spin-spin coupling

Set of Total no. of Identification No. of proton in

equivalence proton in a of different neighboring
protons structure groups carbon (n+1 rule)
Number of signals from NMR spectrum a a b a
No. of signals (Equivalent and non-equivalent protons) b O c H3C CH CH3
H2C C CH3
All of the protons in a molecule which are in OH
c
chemically identical environments will often exhibits 3 signals 3 signals
the same chemical shift and shows one signal in In geometric isomers, stereo-chemically
NMR spectrum at the same value of δ. The protons molecules have equivalence protons.
in this case are said to be chemically equivalent. On a b
the other hand, molecules which have sets of H 3C H Here, b and c is not in
protons which are chemically distinct (have different C C chemically equivalent
Cl H
chemical environments) from one another give rise c
to different absorption signals from each other.
a a
12 11 10 9 8 7 6 5 4 3 2 1 0
a O a H2C CH2 a b c
trans- isomers cis- isomers in
H3C C CH3 H2C CH2 H3C CH2 NH2 in more upfield more upfield
a a
1 signals 1 signals 3 signals
Number of signals from NMR spectrum
a a
Area of signals (Integration of a peak)– b
H3C CH CH3
Not only does each different type of hydrogen give a
distinct peak in the NMR spectrum, but we can also tell Br
the relative numbers of each type of hydrogen by a
process called integration. a
Integration = determination of the area under a peak
 The area under each peak is proportional to the number of b
hydrogens generating that peak.

 The integrated area measured by a ruler are

a : b.
 Divide by the smallest number give us the simplest ratio.
 Note that the integration gives only ratios, not absolute
values for the number of the hydrogen present in the
The 1H NMR spectrum of 2-bromopropane. The -CH3
proton signal at 1.71 𝜹𝜹 is split into a doublet, and the –
CHBr- proton signal at 4.28 𝜹𝜹 is split into a septet. Note
that the distance between peaks — the coupling constant
— is the same in both multiplets. Note also that the two
outer peaks of the septet are small enough to be nearly
missed.

sample.
Number of signals from NMR spectrum
Spin-spin coupling (Signal splitting – n+1 rule) – n+1 rule
 Often a group of hydrogens will appear as a This hydrogen’s peak These hydrogens are
is split by its two split by their single
multiplet rather than as a single peak. This happens
neighbors neighbor
because of magnetic interactions with neighboring
hydrogens with nonequivalent protons on adjacent
carbons and so, is called Spin-Spin splitting.

 The splitting is into one more peak than the H H H H

number of H’s on the adjacent carbon(s), This is C C C C
the “n+1 rule”.
H H
 The relative intensities are in proportion of a
binomial distribution given by Pascal’s Triangle.
Two neighbors One neighbor
n+1 = 3 n+1 = 2
 The set of peaks is a multiplet ( 𝟐𝟐 = 𝒅𝒅𝒅𝒅𝒅𝒅𝒅𝒅𝒅𝒅𝒅𝒅𝒅𝒅, 𝟑𝟑 Triplet Doublet
= 𝒕𝒕𝒕𝒕𝒕𝒕𝒕𝒕𝒕𝒕𝒕𝒕𝒕𝒕, 𝟒𝟒 = 𝒒𝒒𝒒𝒒𝒒𝒒𝒒𝒒𝒒𝒒𝒒𝒒𝒒𝒒, 𝟓𝟓 = 𝒑𝒑𝒑𝒑𝒑𝒑𝒑𝒑𝒑𝒑𝒑𝒑, 𝟔𝟔 = 𝒉𝒉𝒆𝒆𝒆𝒆𝒆𝒆𝒆𝒆𝒆𝒆, 𝟕𝟕
= 𝒉𝒉𝒉𝒉𝒉𝒉𝒉𝒉𝒉𝒉𝒉𝒉 … . .)
Number of signals from NMR spectrum
Spin-spin coupling (Signal splitting – Pascal's triangle) – Singlet (s)
Doublet (d)
Triplet (t)
Quartet (q)
Quintet (quin)
Sixtet (six)
Septet (sept)
Ocetet (oct)
Nonet (non)
Multiplet (m)
Number of signals from NMR spectrum
Spin-spin coupling (Signal splitting - Origin) –

Spin-spin splitting arises because

hydrogen on adjacent carbon atoms
can “sense” one another.
1H NMR—Spin-Spin Splitting
Whenever two (or three) different sets of adjacent protons are equivalent to each other, use the
n + 1 rule to determine the splitting pattern.
Thanks to everyone