Nuclear Magnetic

Resonance (NMR)
Spectroscopy
 Introduction
•  NMR is the most powerful tool available for
organic structure determination and
elucidation.

•  Certain nuclei have spin states of +1/2 and

-1/2

•  This property allow them to be studied by

NMR (NMR active)
2
 Introduction

•  NMR is used to study a wide variety of nuclei:

Ø 1H
Ø 13C
Ø 15N
Ø 19F
Ø 31P

3
 Nuclear Spin
•  A nucleus with an odd atomic number or an odd mass
number has a nuclear spin.

•  The spinning charged nucleus generates a magnetic

field – magnetic dipole

4
 External Magnetic Field
•  When placed in an external field (Bo), spinning
protons act like bar magnets.

5
 External Magnetic Field
•  Therefore the spin states of a nucleus is affected by
an applied magnetic field (Bo)

6
 Two Energy States
•  The magnetic fields of
the spinning nuclei will
align either with the
external field, or
against the field.

•  A photon with the right

amount of energy can
be absorbed and cause
the spinning proton to
flip.

7
 Two Energy States
•  The energy difference between the two spin states depends
on the strength of the magnetic field

8
 Two Energy States
•  The energy difference between the two spin states depends
on the strength of the magnetic field

absorb ΔE
α-spin states β-spin states
release ΔE

Signals detected by NMR

9
 ΔE and Magnet Strength
•  Energy difference is proportional to the magnetic field
strength.

•  ΔE = hν = γ h B0
2π

•  Gyromagnetic ratio, γ, is a constant for each nucleus

(26,753 s-1gauss-1 for H).

•  In a 14,092 gauss field, a 60 MHz photon is required

to flip a proton.
10
 ΔE and Magnet Strength
•  Low energy - radio frequency

ΔE = hν = γ h B0
2π 11
 Magnetic Shielding
•  If ALL protons absorbed the same amount of energy in
a given magnetic field
Ø  not much information could be obtained
Ø  a single NMR absorption will be observed

•  However protons are surrounded by electrons that

shield them from the external field
Ø  Therefore absorption frequency is not the SAME for all H`s

•  Circulating electrons create an induced magnetic field

that opposes the external magnetic field.

12
 Magnetic Shielding
•  ALL nuclei on a molecule is surrounded by
electrons.

•  When magnetic field is applied to the molecule,

electrons moving around the nuclei sets up tiny
magnetic fields of their own.

•  This magnetic field act in opposition to the

applied field.

13
Magnetic Shielding

14
Magnetic Shielding

Y= electronegative atom 15
 Shielded Protons
•  Magnetic field strength must be increased
for a shielded proton to flip at the same
frequency.

16
 Protons in a Molecule
•  Depending on their chemical environment,
protons in a molecule are shielded by
different amounts.

17
 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.
18
Basic NMR Spectrometer

19
NMR Spectrometer
600 MHz NMR Spectrometer

Cost: 2 millions (NZD)

20
The NMR Spectrum

21
 CH3
H3C Si CH3
CH3 Tetramethylsilane (TMS)

•  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.
22
 CH3
H3C Si CH3
CH3 Tetramethylsilane (TMS)

23
 Chemical Shift
•  Measured in parts per million.

•  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. 24

Delta Scale

Chapter 13 25
Location of Signals
•  More electronegative
atoms deshield more and
give larger shift values.

•  Effect decreases with

distance.

•  Additional electronegative
atoms cause increase in
chemical shift.
26
Typical Values

Chapter 13 27
 Typical Values
•  The electrons surrounding a nucleus affect the effective
magnetic field sensed by the nucleus

28
 Factors Affecting Chemical
Shifts
Hybridisation Effect

•  sp3 hybridized H’s - resonate between 0-2 ppm

29
 Factors Affecting Chemical
Shifts
Hybridisation Effect

•  sp2 hybridised H’s - resonate downfield than aliphatic

protons
H
C C 4.5-7 ppm

H
7-8 ppm

H
9-10 ppm
O C
30
 Factors Affecting Chemical
Shifts
Diamagnetic Anisotropy
•  π electrons are less tightly held by the nuclei than σ
electrons; ⇔ more free to move in a magnetic field

This “diamagnetic
anisotropy” is
Diamagnetic anisotropy
commonly used to
- Mean different regions in
rationalize the
space will have different
unusual chemical
magnetic strengths
shifts of protons in
acetylene, aromatic
and antiaromatic
compounds.

•  Causes unusual chemical shifts downfield for hydrogen

31
bonded to carbons that form π bonds (7-8 ppm)
Aromatic Protons, δ7-δ8

Chapter 13 32
Vinyl Protons, δ5-δ6

33 =>
Acetylenic Protons, δ2.5

•  π e in C≡C circulating produce

magnetic field opposing B0.

•  H is shielded by induced magnetic

field - resonate at lower frequency 2-3
ppm

34
Aldehyde Proton, δ9-δ10

Electronegative
oxygen atom

35
Annulene Protons
•  The inner hydrogen atoms are
highly shielded

•  While the outer hydrogen atoms

are highly deshielded

•  Inner protons resonate at -2.99 ppm

(6H)

•  Outer protons resonate at 9.28 ppm

(12H)

36
 O-H and N-H Signals
exchangeable protons
•  Chemical shift depends on concentration.

•  Hydrogen bonding in concentrated

solutions deshield the protons, so signal
is around δ3.5 for N-H and δ4.5 for O-H.

•  Proton exchanges between the molecules

broaden the peak.
37
Carboxylic Acid
Proton, δ10+

38
 Number of Signals
•  Equivalent hydrogens have the same
chemical shift.

Chapter 13 39
 Intensity of Signals
•  The area under each peak is proportional
to the number of protons.
•  Shown by integral trace.

40
 Intensity of Signals

•  2 different types of H’s

•  # of H’s represented on each signal is same 41
 How Many Hydrogens?
•  When the molecular formula is known,
each integral rise can be assigned to a
particular number of hydrogens.

42
 Spin-Spin Splitting
•  Protons in chemically identical environment and
usually magnetically equivalent.

•  Chemically equivalent protons resonate at same

chemical shift.

•  1H NMR tells us the # of different H’s.

43
 Spin-Spin Splitting
•  Nonequivalent protons on adjacent carbons have
magnetic fields that may align with or oppose the
external field.

•  This magnetic coupling causes the proton to

absorb slightly downfield when the external field is
reinforced and slightly upfield when the external
field is opposed.

•  All possibilities exist, so signal is split

44
Spin-Spin Splitting

45
 1,1,2-Tribromoethane
Nonequivalent protons on adjacent carbons.

Chapter 13 46
Doublet: 1 Adjacent Proton

Chapter 13 47
Triplet: 2 Adjacent Protons

Chapter 13 48
 The N + 1 Rule
•  If a signal is split by N equivalent protons,
it is split into N + 1 peaks.

49
 The N + 1 Rule
•  Therefore when a signal is observed in the 1H NMR spectrum,
often it is split into multiple peaks

•  Multiplicity or a splitting patterns results

50
 Range of Magnetic
Coupling
•  Equivalent protons DO NOT split each other.

•  Protons bonded to the same carbon will split each

other only if they are not equivalent.

•  Protons on adjacent carbons normally will couple.

•  Protons separated by four or more bonds will not

couple.
51
Splitting for Ethyl Groups

52
 Splitting for
Isopropyl Groups

=>
53
 Coupling Constants
•  Distance between the peaks of multiplet

•  Measured in Hz

•  Not dependent on strength of the external field

•  Multiplets with the same coupling constants may

come from adjacent groups of protons that split
each other. 54
 Values for
Coupling Constants

Chapter 13 55
 Coupling to more than one
coupling partner

•  If a proton A is coupled to more than one coupling partner

(M and X), its signal will become more complex, if JAM ≠ JAX
•  For each coupling, the n+1 rule should be applied
separately
 a c
H H
C C
Hb Complex Splitting

•  Signals may be split by adjacent protons,

different from each other, with different
coupling constants.

Example
•  Ha of styrene which is split by an adjacent Hb
trans to it (J = 17 Hz) and an adjacent Hc cis
to it (J = 11 Hz). 57
H
a
H
c
Splitting Tree
C C
Hb

58
Spectrum for Styrene

59
 Coupling patterns for disubstituted aromatic
systems

dd
dd 7.6 7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.p8p

td
td

dt t

t
dt 7.8 7.6 7.4 7.2 7.0 6.8pp

“d”

“d”
7.8 7.6 7.4 7.2 7.0 6.8pp

3J = 6 – 10 Hz
AB(ortho)
4J
AB(meta) = 1 – 3 Hz
5J
AB(para) = 0 – 1 Hz (normally not visible)
Stereochemical Nonequivalence
•  Usually, two protons on the same C are
equivalent and do not split each other.

•  If the replacement of each of the protons of a -

CH2 group with an imaginary “Z” gives
stereoisomers, then the protons are non-
equivalent and will split each other.

•  They are called diastereotopic protons

61
Diastereotopic Protons

62
Some Nonequivalent
Protons
a c c
H H H OHa
C C
Hb dH Hb

CH3
H Cl
aH Hb
Cl

63
 Time Dependence
•  Molecules are tumbling relative to the magnetic
field, so NMR is an averaged spectrum of all the
orientations.

•  Axial and equatorial protons on cyclohexane

interconvert so rapidly that they give a single
signal.

•  Proton transfers for OH and NH may occur so

quickly that the proton is not split by adjacent
protons in the molecule.
64
Hydroxyl
Proton
•  Ultrapure samples of
ethanol show
splitting.

•  Ethanol with a small

amount of acidic or
basic impurities will
not show splitting.

65
 N-H Proton
•  Moderate rate of exchange.
•  Peak may be broad.

66
 Identifying the O-H
or N-H Peak
•  Chemical shift will depend on concentration and
solvent.

•  To verify that a particular peak is due to O-H or N-H,

shake the sample with D2O

•  Deuterium will exchange with the O-H or N-H

protons.

•  On a second NMR spectrum the peak will be absent,

67
or much less intense.
The basic of 13C NMR Spectroscopy
•  Most of the theories about 1H NMR spectroscopy also
applies to 13C NMR, although there are several important
differences.

1.
•  The magnetic moment (hence NMR signals) of a 13C
nucleus is much weaker than that of a proton.

•  The resonance frequencies of 13C nuclei are lower than

those of protons in the same applied field (e.g. in a 7.05 T
instrument, protons resonate at about 300 MHz, while
carbons resonate at ~75 MHz).

68
The basic of 13C NMR Spectroscopy
1.
•  This is fortunate, as it allows us to look at 13C signals using
a completely separate 'window’ of radio frequencies.

2.
•  Since 13C nucleus accounts for only 1% of carbon atoms in
nature and has weaker NMR signal,

•  Therefore more sample is required and often the data from

hundreds of scans must be averaged in order to bring the
signal-to-noise ratio down to acceptable levels.

69
The basic of 13C NMR Spectroscopy
3.
•  Chemical shifts for 13C nuclei in organic molecule s are
spread out over a much wider range than for protons (0 to
220 ppm for 13C compared to 0 to 14 ppm for protons)

•  This is also fortunate, because it means that the signal from

each carbon in a compound can almost always be seen as
a distinct peak, without the overlapping that often plagues
1H NMR spectra.

70
The basic of 13C NMR Spectroscopy
4.
•  Unlike 1H NMR signals, the area under a 13C-NMR signal
cannot be used to determine the number of carbons to
which it corresponds.

•  This is because the signals for some types of carbons are

inherently weaker than for other types (e.g. peaks
corresponding to carbonyl carbons are much smaller than
those for methyl or methylene (CH2) peaks).

71
The basic of 13C NMR Spectroscopy
5.

•  However, just like in 1H NMR, the standard used in 13C

NMR experiments to define the 0 ppm point is
tetramethylsilane (TMS)

•  Although of course in 13C NMR it is the signal from the four

equivalent carbons in TMS that serves as the standard.

72
 Carbon-13
•  12C has no magnetic spin

•  13C has a magnetic spin, but is only 1% of the

carbon in a sample

•  The gyromagnetic ratio of 13C is one-fourth of

that of 1H.

•  Signals are weak, getting lost in noise.

•  Hundreds of spectra are taken, averaged. 73

13C NMR Spectrum Methyl Acetate

•  The 13C NMR spectrum of methyl acetate shows three

peaks, one for each of the three chemically distinct carbon
atoms in the molecule.

•  What are the other small signals?

74
 Fourier Transform NMR
•  Nuclei in a magnetic field are given a radio-frequency
pulse close to their resonance frequency.

•  The nuclei absorb energy and precess (spin) like little

tops.

•  A complex signal is produced, then decays as the

nuclei lose energy.

•  Free induction decay (FID) is converted to spectrum.

75
 Fourier Transform NMR
•  Numerous individual runs are added together and
averaged such that random background noise cancels
to zero

•  The NMR signals are enhanced, substantially

increasing sensitivity.

76
 Fourier Transform NMR
•  A single run showing background noise

•  An average of 200 runs

77
Hydrogen and Carbon
Chemical Shifts

Chapter 13 78
=>
Carbon Chemical Shifts
Combined 13C
and 1H Spectra

80
 Differences in
13C Technique

•  Resonance frequency is ~ one-fourth,

15.1 MHz instead of 60 MHz.

•  Peak areas are not proportional to

number of carbons.

•  Carbon atoms with more hydrogens

absorb more strongly. 81
 Spin-Spin Splitting

•  Because of the low natural abundance of 13C nuclei,

•  It is unlikely that a 13C would be adjacent to another

13C, so splitting by carbon is negligible.

•  There is, however, heteronuclear coupling between

13C and 1H

•  13Cwill magnetically couple with attached protons

and adjacent protons
82
 Spin-Spin Splitting
•  Carbon-proton coupling constants are very large (on the
order of 100-250 Hz)

•  These complex splitting patterns are difficult to interpret,

•  A technique called broadband decoupling, which essentially

'turns off’ 13C-1H coupling resulting in a spectrum in which all
carbon signals are singlets, is mostly used.

83
 Proton Spin Decoupling
•  To simplify the spectrum, protons are
continuously irradiated with “noise,” so
they are rapidly flipping.

•  The carbon nuclei see an average of all

the possible proton spin states.

•  Thus, each different kind of carbon gives

a single, unsplit peak. 84
Off-Resonance Decoupling
•  13C nuclei are split only by the protons
attached directly to them.

•  The N + 1 rule applies: a carbon with N

number of protons gives a signal with
N + 1 peaks.

85
 Interpreting 13C NMR
•  The number of different signals indicates the
number of different kinds of carbon.

•  The location (chemical shift) indicates the type of

functional group.

•  The peak area indicates the numbers of carbons

(if integrated).

•  The splitting pattern of off-resonance decoupled

spectrum indicates the number of protons
86
attached to the carbon.
Two 13C NMR Spectra

87
 MRI
•  Magnetic resonance imaging, noninvasive

•  “Nuclear” is omitted because of public’s fear that

it would be radioactive.

•  Only protons in one plane can be in resonance at

one time.

•  Computer puts together “slices” to get 3D.

•  Tumors readily detected. 88