Nuclear Magnetic

Resonance Spectroscopy
 Nuclear Magnetic Resonance
Spectroscopy

magnetic field quantizes nuclear energy

levels, where electromagnetic radiation
can be absorbed to induce transitions
(radio frequency is of the proper energy)
 Magnetic Properties
Nucleus: 1H
Magnetogyric ratio: 2.68 x 108 radian/T·s
Isotopic Abundance: 99.98%
Relative Sensitivity: 1.00
Absorption Frequency: 200 MHz
at a field strength of 4.69 T
 Magnetic Properties
Nucleus: 13C
Magnetogyric ratio: 6.73 x 107 radian/T·s
Isotopic Abundance: 1.11%
Relative Sensitivity: 0.016
Absorption Frequency: 50.2 MHz
at a field strength of 4.69 T
 Magnetic Properties
Nucleus: 19F
Magnetogyric ratio: 2.52 x 108 radian/T·s
Isotopic Abundance: 100.0%
Relative Sensitivity: 0.83
Absorption Frequency: 188 MHz
at a field strength of 4.69 T
 Magnetic Properties
Nucleus: 31P
Magnetogyric ratio: 1.08 x 108 radian/T·s
Isotopic Abundance: 100.0%
Relative Sensitivity: 0.066
Absorption Frequency: 81.0 MHz
at a field strength of 4.69 T
Magnetic Moments and Energy Levels

Magnetic
moments and
energy levels for a
nucleus with a
spin quantum
number of + 1/2
 Precession in Magnetic Field

“Precession of a
rotating particle in a
magnetic field.”
 Procession of a Magnetic Moment on
Application of a Steady Magnetic Field
Nuclear Spin Quantum Number, I

2I + 1 states
0, 1/2, 1, 3/2, etc 11/2
identical energies in absence of applied
magnetic field
Nuclear Spin Quantum Number, I
No. of No. of Value of
Protons Neutrons I Examples
Even Even 0 12C, 16O, 32S

Odd Even 1/2 1H, 19F, 31P

3/2 11B, 79Br

Even Odd 1/2 13C

3/2 127I

Odd Odd 1 2H, 14N

 Other NMR Nuclei
spin 1/2 nuclei
• Fe-57, Ag-107, Ag-109, Sn-117, Sn-119,
W-183, Pt-195, Hg-199, and Pb-207
Quadrupolar nuclei with spins from 1 to 9/2
• nitrogen-14 I = 1 lithium-7 I = 3/2
boron-10 I = 3 oxygen-17 I = 5/2
cobalt-59 I = 7/2 indium-115 I = 9/2.
 Nuclear
orientation
and energy
levels of
nuclei in a
magnetic field
for different
spin numbers.
 Magnetic Quantum Number

m => magnetic quantum number

...
values => I, I - 1, , -I
 Resonance Energy
mh
E= - -------- Bo
2
where => magnetogyric ratio
m => magnetic quantum number
h => Planck's constant
Bo => strength of external field
 Resonance Energy
when m = +1/2

(+1/2)h h
E+1/2 = - ------------ Bo = - ----- Bo
2 4

-
( 1/2)h h
E-1/2 = - ------------ Bo = ----- Bo
2 4
 Resonance Energy

-
E = Ehigher stste Elower state = E-1/2 -E +1/2

E = (hBo/ 4 - (- h B / 4

o

h
E = h = -----Bo thus =(Bo )/2
2
 NMR frequency in MHz
Isotope Magnetic
Moment,  14.09 kG 23.49 kG
1H 2.79268 60.000 100.000
13C 0.70220 15.086 25.147
19F 2.6273 56.444 94.087
31P 1.1305 24.288 40.485
 NMR field values in kG
Isotope Magnetic
Moment,  60 MHz 100 MHz
1H 2.79268 14.092 23.49
13C 0.70220 56.05 93.407
19F 2.6273 14.98 24.965
31P 1.1305 34.81 58.019
 NMR
spectra of
acrylonitrile
at 60, 100,
and
220 MHz
EXAMPLE: Many NMR instruments
employ a magnet that provides a field
strength of 14,092 G (1.41 T). At what
frequency would the 13C nucleus absorb in
such a field?
=(Bo )/2
where 6.73 x 107
= (6.73 x 107  1.41 )/2
= 15.1 MHz
Distribution of Particles Between
Magnetic Quantum States

- -
Nj/No = exp ( E/kT) = exp ( h/kT)
Example: Calculate the relative number of 13C
atoms in the higher and lower magnetic states
when a sample is placed in a 1.41 T field at
20oC.
Nj/No = exp (-E/kT) = exp (-h/kT)
where = 15.1 MHz
Nj/No=exp(- ((6.63e-34*15.1e6)/(1.38e-23*293)))
= 0.999999752
for 106 in higher state
No = 106/ 0.999999752 = 1,000,000.24
0.24 ppm enrichment
Example: Calculate the relative number of
13C atoms in the higher and lower magnetic
states when a sample is placed in a 4.69 T
field at 20oC.
- -
Nj/No = exp ( E/kT) = exp ( h/kT)
where = 50.2 MHz
Nj/No=exp(- ((6.63e-34*50.2e6)/(1.38e-23*293)))
= 0.99999177
for 106 in higher state
No = 106/ 0.999999177 = 1,000,008
8 ppm enrichment
Example: Calculate the relative number of
13C atoms in the higher and lower magnetic
states when a sample is placed in a 20.5 T
field at 20oC.
- -
Nj/No = exp ( E/kT) = exp ( h/kT)
where = 220 MHz
Nj/No=exp(- ((6.63e-34*220e6)/(1.38e-23*293)))
= 0.999963927049
for 106 in higher state
No = 106/ 0.999963927049 = 1,000,036
36 ppm enrichment
Example: Calculate the relative number of
13C atoms in the higher and lower magnetic
states when a sample is placed in a 89 T field
at 20oC.
- -
Nj/No = exp ( E/kT) = exp ( h/kT)
where = 900 MHz
Nj/No=exp(- ((6.63e-34*900e6)/(1.38e-23*293)))
= 0.999852
for 106 in higher state
No = 106/ 0.999852 = 1,000,148
148 ppm enrichment
 Environmental Effects
1. number of signals
2. position of signal - chemical shift
3. intensity of signal
4. splitting of signals
 Number of Signals

“(a.) Partial spectrum of

methyl-2,3,4-tri-O-
benzoyl--L-
lyxopyranoside, 100 MHz
CDCl3. (b.) H2 and H3
decoupled (c.) H4
decoupled. Note that
there are two H5 protons.”
Position of Signal Chemical Shift
“Absorption positions of protons in various
structural environments.”
 Splitting of Signal
“The effect of the methylene protons in
ehtanol on the resonance of the methyl
protons.”
 Splitting of Signals
No. Eq. Relative
Protons Multiplicity Peak Areas
0 1 1
1 2 1 1
2 3 1 2 1
3 4 1 3 3 1
4 5 1 4 6 4 1
5 6 1 5 10 10 5 1
6 7 1 6 15 20 15 6 1
 Applications of Proton NMR
identification of unknowns
20 ppm range of chemical shifts
almost solvent independent
Continuous-Wave NMR
 Continuous-Wave NMR
“Block diagram of Continuous-Wave NMR.”
 Instrumentation - CW-NMR

R-F transmitter
R-F receiver & detector
sweep generator => sweep coil
sample
magnet
recorder
 Instrumentation CW-NMR

R-F transmitter
– irradiate sample with resonant radio frequency
– less than 1 W output but reproducible to 1 part
in 109
 Instrumentation CW-NMR

R-F receiver & detector

– detect the radio frequency
– amplify 105 times
– integrator
 Instrumentation CW-NMR

sweep generator => sweep coil

– modify the applied magnetic field
– “scans” through slight (ppm) changes in
magnetic field which produces variations in
position of resonant frequency
– modifies magnetic field to give spectrum
 Instrumentation CW-NMR

sample
– sample suspended between pole faces of
magnet
– sample is spun
• air driven turbine - average out effects of in
homogeneities in the field
– 5 mm O.D. tube
 Instrumentation CW-NMR

magnet
– causes the nuclear spin states to split
– allows sample to absorb energy equal to the
difference between spin states
– most expensive component
– must be reproducible & homogeneous
 Magnets
type of magnet max. field strength
permanent 14,092 G
conventional electromagnet 23,490
superconducting solenoids <169,000
 Magnets
type of magnet max. field strength
permanent 1.41 T
conventional electromagnet 2.35
superconducting solenoids <16.9
 Instrumentation
recorder
– displays the spectra
 Sample Handling
– solution
– solvents should be free of hydrogen
• deuterated solvents used where necessary
– should contain standard
• most commonly tetramethyl silane (TMS)
• TMS signal position set at 0 ppm
• other peaks relative to TMS
FT-NMR
 FT-NMR

Anasazi Instruments EFT-60

 FT-NMR
FT advantage
multiplex advantage
improved magnets
 FT-NMR
Pulsed Excitation
– short pulse of strong R-F signal irradiates
sample, 10:s
– sample emits radiation, free induction decay
(FID)
 FT-NMR

time domain spectra

instead of
frequency domain
"(a) FID signal for
13C in cyclohexane.

(b) Fourier
transform of (a)."
 FT-NMR
Free Induction Decay
– all of the nuclei will re-emit RF radiation at
their respective resonance frequencies
– interference pattern in the resulting RF
emission versus time
– frequencies are extracted from the FID by a
Fourier transform of the time-based data.
 Spin Decoupling
"Effect of spin decoupling on the NMR
spectrum of nicotine dissolved in CDCl3.
Curve A, the entire spectrum. Curve B,
expanded spectrum for the four protons
on the pyridine ring. Curve C, spectrum
for protons (a) and (b) when decoupled
from (d) and (c) by irradiation with a
second beam that has a frequency
corresponding to about 8.6 ppm."
 Chemical Shift Reagents
– disperses absorption peaks over a larger
frequency range
– usually compounds of europium and
praseodymium
 Chemical Shift Reagents
"Effect of
Pr(DPM)3 on
NMR spectrum of
styrene oxide.
Spectrum A, in
the absence of the
reagent.
Spectrum B, in
the presence of
the reagent."
Components of FT Spectrometers

"Block diagram
of a Fourier
transform NMR
spectrometer."
 Magnet
– in the past, electromagnets or superconducting
solenoids
– today, permanent magnets or superconducting
solenoids
Locking the Magnetic Field

– field/frequency lock system

– reference nucleus is continuously irradiated and
monitored at a frequency corresponding to its
resonance maximum at the rated field strength
of the magnet
– provided by deuterium in the solvent, with
second transmitter coil set to the frequency for
deuterium
 Shimming
– coils which provide some changes in the
magnetic field by carefully controlling the
current flowing through the coils
– used to correct for inhomogeneities of magnet
 Sample Spinning
– sample is spun along its longitudinal axis to
counteract some of the field inhomogeneities
– "A minor disadvantage of spinning is that the
magnetic field is modulated at the spinning
frequency, which may lead to sidebands on
each side of absorption peaks?"
 Sample Probe
1.holds sample in the magnetic field
2.spins the sample
3.houses the coil or coils that permit
excitation and detection of the NMR
signal and preamp
4.coil for locking
5.coil for decoupling
6.variable temperature capability
Transmitter/Receiver Coils

– early instruments, separate coils, at right angles

to each other
– FT-NMRs, normally same coil
 Pulse Generator
1. a continuous crystal oscillator
2. a gate to switch the pulse on and off
3. a power amplifier to amplify the pulse to
perhaps 50 - 100 W
 Receiver System
– nanovolt - microvolt signal amplified to 10 V
 Detector and Data
Processing System
– high-frequency radio signal converted to audio-
frequency signal to digitized
– carrier signal with superimposed NMR signal,
10 ppm PMR spectrum at 270 MHz would have
a frequency range of 2700 Hz
– subtract the carrier signal from NMR signal
 Sampling the Audio Signal
digitize signal, must sample signal at least
twice per cycle
 Single-Channel Detection
disadvantage, does not know if difference
between carrier signal and NMR signal is
positive or negative, can produce a false
signal
Quadrature Detection Systems
– NMR signal is split and fed to two identical
detectors
• in one detector the signal is treated in the normal
manner, carrier signal subtracted from NMR
signal for digitizing
• in the second detector the carrier signal is altered
900 before subtraction
Quadrature Detection Systems

– the two audio signal are the same except for

phase, the first produces a cosine wave while
the second produces a sine wave
– the two signals are digitized, transformed
separately into frequency domain signals
and combined to produce the spectrum
– folded signal disappears because it is of
opposite sign in the cosine and sine wave
forms
Quadrature Detection Systems
"A quadrature detector system."
 Quadrature Detection Systems
"Fourier transform of (A) a cosine and (B) a sine time
domain signal from the quadrature detector.
(C) Combination of A and B giving and NMR
spectrum."
 2D FT-NMR
“Illustration of the use
of the two-
dimensional spectrum
(a) to identify the 13C
peaks in a one-
dimensional spectrum
(b).”
270MHz H-1 FT-NMR
270MHz C-13 FT-NMR
270MHz 2D FT-NMR