Common types of NMR experiments:

1-H NMR

a. Experiment – High field proton NMR (400MHz).

single-pulse experiment.
Proton NMR
b. Spectral Interpretation

i. Number of multiplets gives the different H-environments

One Dimensional H-NMR ii. Splitting patterns indicate number of protons on ‘adjacent Cs’
iii. Integration, indicates relative number of types of protons.
iv. Chemical shift (ppm), indicates chemical environment.

1 2

Free rotation about sigma bonds

A H nucleus that is surrounded by higher electron density
Cl F make the nuclei equivalent on the
will generally come into resonance at lower frequency than a
nucleus surrounded by less electron density. NMR time scale.
S
Thus the number of ‘multiplets’ gives the number of different higher electron density
H-environments in the molecule. lesser electron density

Low field High field

The ‘mean position’ of resonance of a group of nuclei is termed High frequency Low frequency
the chemical shift, δ, (ppm). δ indicates chemical environment.

A chemical shift a measure of the resonance frequency of a

particular type of nucleus compared to that of a standard
molecule, TMS in 1H-NMR, scaled to the frequency of the
spectrometer and reported as parts per million, ppm.

3 4

Cl

Ratio of the integrated of peaks (peak areas/heights) indicates Cl F

relative number of protons in different environs.
S
Peak (multiplet) area ∝ nH generating the signal, however
area peaks are not perfect.

5 6
Hyperfine Splitting; (J-coupling/ scalar coupling)

NMR signals split into multiple peaks when molecules contain

non-equivalent hydrogen atoms that are separated by covalent
Cl
(usually no more than three bonds for saturated compounds).

The multiplets results from the spin-spin coupling

between nuclei, an interaction in which nuclear spins of
non-equivalent adjacent atoms influence each other.

Signal splitting allows the determination of how different H-

carrying atoms are connected in a molecule, because atoms
on adjacent ‘functional’ groups can generally split each other.

The general rule is that a signal will be split into (n+1) peaks
if there are n equivalent (or nearly equivalent) H atoms three
bonds away. 7 8

H H
Cl CH3
c
H H
a b
Equivalent protons – protons that are having the same
chemical shift and spin coupling.

9 10

H
H Cl
H3C CH3 H H
Cl HH HH

Splitting (patterns) are indicative of the equivalence (nearly) or

nonequivalence of the adjacent H (environs).

11 12
 Cl I Cl I

HX Cl I Cl I
HA
H H H H HX
X A X A
Cl I Cl I stem
Cl I Cl I Coupling Constant
JXA lines
H H
↓ ↓ H H ↓
X A X A
Cl I Cl I ↓HA ↑ HA
Cl I Cl I
H H ↑ ↑ H H ↑
X A X A The coupling constant is independent of the applied field.

13 14

HX HA
HX HA
↓ ↑ ↓ ↑
Tree diagram

JXA JAX
Coupling Constant
Hyperfine splitting arises due to the coupling (transmission
of the spin state information) of the adjacent magnetic
nuclei via bonding electrons. HX feels the spin orientations of HA.

This is a through bond coupling process.

JAX=JXA

The coupling constant is independent of the applied field.15 16

Cl I

Cl I

H H

In vicinal coupling is the dihedral angle a between the adjacent

C-H bonds and whether or not it is fixed is important. Coupling is
highest when the angle 0° and 180°, and is lowest at 90°.

Bonds that rotate rapidly at room temperature do not have a

fixed angle between adjacent C-H bonds, so an average angle
and an average coupling is observed.

This latter concept is important for the interpretation of 1H-NMR

spectra for alkanes and other flexible molecules.

17 18
 Ha

Note the splitting constant J; for ‘groups’ of atoms with spins

coupled, J values are the same!!

#peaks (2) (3) (4)

19 20

In a first-order spectrum where chemical shift difference is

much larger than their coupling constant (nuclei A and nuclei X),
The splitting arises because there are n+1 different possible
∆δA-Xνref > 6 JA-X) the interpretation of splitting patterns is
spin state combinations of n spins, aligned and against the
straightforward, multiplets are well separated.
external magnetic field arising from n equivalent protons
The multiplicity of a multiplet is given by the number of
The probability of a molecule having a given set of spins is
equivalent neighboring equivalent atoms, n plus 1, (n+1 rule).
proportional to the number of possible spin alignments giving
rise to that spin state.
Equivalent nuclei do not interact with each other.
Peaks in the multiplet pattern conforms to the coefficients
The coupling constant is independent on the applied field.
of the Binomial expansion (Pascal triangle - first order
spectra).
Multiplets can be easily distinguished from closely spaced
chemical shifts.

Peaks in the multiplet pattern conforms to the coefficients

21 of the Pascal triangle. 22

Splitting by more than one non-equivalent atoms Splitting by more than one non-equivalent atoms

To predict the splitting pattern use a tree diagram appplying To predict the splitting pattern use a tree diagram appplying
different couplings applied sequentially. different couplings applied sequentially.

JAB>JBC JAB>JBC

General case, a signal will be split into (n + 1)(m + 1) peaks General case, a signal will be split into (n + 1)(m + 1) peaks
for an H atom that is coupled to a set of n H atoms with J for an H atom that is coupled to a set of n H atoms with J
value, and to another set of m H atoms with another J. value, and to another set of m H atoms with another J.
23 24
 Complex splitting pattern is determined by the J values

JAB>JBC (n + 1)(m + 1)

Note the splitting constant J; for ‘groups’ of atoms that are

spin coupled, J values are the same!!
25 (n + 1)(m + 1) (n + 1) 26

27 28

Analysis of first order spectral clusters

d-q-d

Mark the peak

Examine thepositions andstarting
area ratios note the area
from anratios, settingthe
end. Notice thearea
peaks
Draw ratio patterns.
vertical areas
lines ofCompare
the at to
same the
sizeends
the to 1 triangle
Pascal
to indicate values.
the peak positions
The ratio values ordain the type of multiplet at this level.
Determine the splitting at this (last) stage. (doublet here)

a-e called levels Reconstruct peak positions and area ratios before
Repeat the procedure until the process
this last coupling.end up in a single peak;
29
i.e. the peak position without hyperfine splitting. 30
 d-q-d

A A
H H
A H
A H
H H

A = ?; not a H

31 32

t-t Equivalent protons – chemical shift and magnetic (spin

coupling).

Chemical Shift Equivalence

Symmetry makes atoms equivalent. Atoms or groups in

identical physical locations relative to a plane of symmetry
or center of symmetry in a molecule are equivalent.

Rapid processes makes atoms equivalent as well.

Area ratio ⇒ Non Binomial

Overlap possible
1:2 ..⇒ 1:2:1?
33 34

H
H Cl
H3C CH3 H H
Cl HH HH

C2 σ i

Diastereoscopic – no σ thro’ CH2; nonequivalent

35 36
Rapid Interconversions Magnetic Equivalence (Spin-Coupling equivalence):

a. Keto-enol tautomerism Applicable (to be considered) for chemical shift equivalent

b. Around partial double bonds protons only. The chemical shift equivalent protons must have
c. Rings the same spin couplings to other nuclei to be magnetically
d. Rotation around sigma bonds equivalent.
JAX ≠ JA’X
JAX’≠JA’X’

Non-equivalent protons at o,p positions

38
Note spectral symmetry

Splitting (patterns) are indicative of the equivalence (nearly) or

nonequivalence of the adjacent H (environs).

39 40

Geminal coupling

Non-conformation to the ‘n+1’ rule

Unsymmetrical molecules with restricted bond rotation such as
alkenes or rings, as well as molecules for which stereochemistry
is important (where equivalence do not exist).
Note the splitting pattern for such systems are complex.
The dihedral angle between C-H bonds determines the extent of
coupling (J). Therefore the bond rotation is a key parameter.
Where free rotation about the C-C σ bonds exist, H - C bonds
are equivalent (e.g. -CH2- and -CH3) due to the rapid bond
rotation (exception -CH2- is adjacent to a stereo-center).

Where there is restricted bond rotation (alkene or cyclic) H’s

bonded to same C atom may not be equivalent, especially if the
molecule is not symmetrical, the nonequivalent H nuclei on the
same C atom will couple to each other and generating splitting
41
of the peaks - geminal coupling. 42
 AMX, ABX, ABC

a, b, c nonequivalent

43 44

45 46

The tree diagram is useful to identify the spin coupled atoms Strongly Coupled (Second and Higher order) Spectra

In these spectra the simple rules used to construct multiplets

(area ratios) are no longer valid. This is because the chemical
shift differences between the spins (nuclei A and nuclei B)
are small compared to the couplings ∆δA-Bνref < 8 JA-B.
Peak splitting can range less to more than expected and
unusual relative peak intensities arise; second-order spectra.

The overall effect of strong coupling is the roofing/tilting of

multiplets.

Jab a
Jac Jde
d e
b c
Jbc
47 48
 Exchangeable H atoms

-OH, -SH, -NH

1. OH, NH protons are exchangeable (fast in NMR scale – no

The degree of leaning depends
on proximity of the chemical
shifts of the absorption, and
The strength of the coupling.
Closer ν values and larger
J values leads to greater leaning
of the ‘roof’.
coupling, single peak or merged into background). The
acidic H signal broadens, do not undergo coupling. H-N
would couple to (3J) adjacent protons.
2. OH, SH, NH, H- bonded
a. intermolecular (δ - solvent, concentration, temperature
dependent)
b. intramolecular

3. 14N(I=1, nuclear quadrupole interacts, lowers lifetime of H

excited states, single broader peak or merged into bkg.)
49 50

In solvents where
exchange could
not occur.

51 52

O OH

Detection of acidic (exchangeable) protons P O HOD

O O
CH3
impurity
In D2O
Acidic protons can be exchanged with deuterium ions.

Mixing a compound with D2O achieves this task. The H-NMR

spectrum of the exchanged product would be absent of the
acidic H peak. In CDCl3

The exchanged product is HOD, the H of which appears at

4.7ppm as a single peak.

Solvent Effect: Change of solvent generally changes chemical

shifts slightly due to solvent-solute interactions changing the
electronic environments of atoms.

53 54
Long Range Coupling

nJ; n>3 occur in rigid structures; alkenes, alkynes,

aromatics, stained rings.

W conformation of four sigma bonds.

Appendix F for coupling constants.

55 56

H
Cl O O
P O
Cl O
O
Areas under multiplets

In principle the area under the multiplets are proportional

to the number of protons generating the multiplets. (2)

(3)
(1) (1) (2)

57 58

1-D Experiment
Integration and T1
Usual H-NMR experiment is a single channel experiment
repeated; a single pulse experiment, repeated.

Protons are excited via irradiation (Channel 1) and the same

channel is used to observe the FID of the signal.

FID is then Fourier Transformed to obtain the NMR spectrum.

θ=π/2
Note the FID decay
characterized by a time
constant that is related to
the rate of re- equilibration

p1

59 60
d1 AQ d1
 θ=π/2 θ=π/2

T2 = spin-spin relaxation time T2 = spin-spin relaxation time

constant (transverse) constant (transverse)
T1 = spin-lattice relaxation time T1 = spin-lattice relaxation time
constant (longitudinal) constant (longitudinal)
p1 p1

d1 AQ d1 d1 > 5T1 d1 AQ d1 d1 > 5T1

Z Equilibrium, M0 Z After pulse.

ω
A collection of spins in a B Y T2 Y
attains an equilibrium state
creating a net macroscopic
magnetic moment, M0 X X
61 62

θ=π/2 θ=π/2

T2 = spin-spin relaxation time T2 = spin-spin relaxation time

constant (transverse) constant (transverse)
T1 = spin-lattice relaxation time T1 = spin-lattice relaxation time
constant (longitudinal) constant (longitudinal)
p1 p1

d1 AQ d1 d1 > 5T1 d1 AQ d1 d1 > 5T1

Z Z

ω ω
T2 Y T2 Y
T1 T1

X X
63 64

Signal intensity given by the integration of peaks is proportional

to M0’s the number of a given type of atoms resonating at it’s The ‘decay’ of excited nuclei restores the system to equilibrium.
frequency. It takes time to restore, ‘relaxation time’, T1.

However the above fact is dependent on how well the It is imperative that all nuclei relaxes back to equilibrium.
experiment parameters are set up. However every type of nuclei do not decay at the same rate.

Pulse experiments (FT-NMR) are repeated, single pulse
experiments, where the nuclei are ‘excited’ and their decay is
followed (during which time an FID is captured) after each pulse.
All FIDs from the repeated application of a series of pulses
(scans) are summed.
Therefore, if the ‘next’ pulse in the multi-pulse protocol is applied
before complete relaxation; without giving sufficient delay time
(5T1) between pulses, the signal size in the transformed
spectrum is not proportional to the number of nuclei slow
decaying nuclei (usually less).

The summed FID is then transformed into the frequency domain

to generate the traditional spectrum.
65 66
Selective Spin Decoupling: Double Resonance
Decoupling is done as a two channel experiment.
Channel 1: Excite and observe protons (pulse)
Channel 2: Excite the nucleus to be decoupled (CW)
Heteronuclear decoupling: different types of atoms involved
(observed atom and decoupled atom type different)
Homonuclear decoupling: same types of atoms involved
(observed atom and decoupled atom type same)
If the spin states of the nucleus/nuclei adjacent to a certain
nucleus is/are flipped between their two spin states very fast,
equalizing the populations then that particular nucleus would
not be able to distinguish (feel) the spin states of the fast spin
flipping neighbors.
The resonance peak appears as if the neighbor ‘do not exist’.
That is, the particular nucleus gets decoupled from the
neighbors.
The result is the elimination of scalar (J) coupling and it results
in the merging of the otherwise spin coupled ‘peaks’.

Further the peaks of the decoupled nuclei vanishes.

67 68

Homonuclear decoupling
π/2
O OH -CH3 protons
Channel 1 P O
irradiated
O
1H observe O
i.e. methyl Hs decoupled
CH3

p1

d1 AQ

Channel 2
selective
irradiation

-CH2-
69 70

Heteronuclear decoupling
NOE and through space interactions
31P irradiated
O OH
P O i.e. P decoupled Scalar (J) coupling arises due to through bond interactions
O O of nuclei in close proximity.
CH3

Another type of interaction is dipolar coupling which occurs

through space and is dependent on the direct distance between
nuclei. This interaction manifests in decoupling experiments.
1
NOE Effect α
r6
The excess energy provided to the molecule via the Channel 2
(much larger than in double resonance experiment, inverts
populations) finds a ‘way’ to dissipate via dipolar coupling to
the ‘nearby’ nuclei. The excess energy shifts population of
spin states from the ‘equilibrium states’. Thus sets up the
-CH- -CH2- 71
need to reestablish the populations. 72
 Methine H Cl

This in turn increases the population of protons low energy

spin states in close proximity, usually ~5 Angstroms from H3C O
the decoupled nucleus. O
P O
O
O

H3C

Cl
74

Cl

H 3C O
O
P O
O
O

H 3C

Cl

Normal spectrum
methine H
decoupled

Selective NOE
A dispersion like output due
to in homogeneity in B
NOE Difference

75

In NOE difference experiments, a 1H is selectively

pre-irradiated until saturation is achieved. During the
pre-irradiation period, NOE buildup occurs at other 1H nuclei
close in space.

Then a 90o pulse is applied, 90o pulse then creates an

observable magnetization, which is detected during the
acquisition period.
π/2

Channel 1
1H observe
CW
d1 p1

AQ
78
 The final nOe difference spectra are displayed as the
difference between a spectrum recorded with on-resonance
pre-irradiation and a reference spectrum that has all proton
absorptions at the same conditions but off resonance.

79 80

Nuclear Overhauser Effect:

Consider a two channel experiment where one channel excites

a spin I and the other saturates a spin S.

Assume the spins I and S are close enough to have a

dipole-dipole coupling (the through-space interaction) and there
is no spin-spin coupling (i.e. scalar coupling) among I and S.

This experiment leads to the enhancement of the signal from

spin I. This phenomenon is known as the Nuclear Overhauser
Effect (NOE).

This enhancement is due to the change in relative populations

of the energy levels brought about by the saturation of S spins
and the subsequent relaxation processes.

ββ
W1C
βα
W2 W1H

W1H W0
αβ
W1C
αα

H 13CO2- W2:W1:W0=12:3:2 H 13CO2- W2:W1:W0=12:3:2

I S I S
 Note:
The NOE effect occurs through space and is operational within
NMR signals with high multiplicities (> 5) may have very low
a ~5A sphere centered around the irradiated nuclei
intensities and may be less than the level in the output.
(“NOE range”).
E.g. (CH3)2CHCH2OH, multiplicity = 21.
It does not enhance resonance signals away from “range” from
Multiplet overlapping may give complex peak ‘splitting’.
the irradiated ‘centre’.
"Long-range"-couplings (> 3 bond) may be stronger (π bonds).
NOE allows determination of the proximity of the nuclei to the
E.g. CH3COOCH2C=CCH2OH.
irradiated ones, thus is an excellent technique to ascertain
the inter-proton distances/stereochemistry of molecules.
Higher order spectra. Relative intensities of peaks of multiplets
changes extensively; usual patterns not followed.

Spin-spin couplings between protons and other magnetically

active nuclei (e.g., 31P, 13C or 19F) leads to additional peak
85
splitting. 86

Cl

H3C O
O
P O
O
H3C O

Cl
87