CHEM 430 NMR Spectroscopy Part 1 2014

Introduction to Spectroscopy
Spectroscopy is the study of the interaction of matter with the

electromagnetic (EM) spectrum
1. EM radiation displays the properties of both particles and waves
2. This packet of wave and particle properties is called a photon
The term photon is implied to mean a small, massless particle that

contains a small wave-packet of EM radiation/light
3. The energy E component of a photon is proportional to the frequency

n
E = hn
The constant of proportionality is Planks constant, h
CHEM 430 NMR Spectroscopy 2

4. Because the speed of light (c ) is constant, the frequency (n)

(number of cycles of the wave per second) can complete in the
same time, must be inversely proportional to how long the
oscillation is, or wavelength (l):
c
___ hc
n= ___
D E = hn =
l l
5. Amplitude describes the wave height, or strength of the oscillation
6. Because the atomic particles in matter also exhibit wave and

particle properties (though opposite in how much) EM radiation
can interact with matter in two ways:
Collision particle-to-particle energy is lost as heat
and movement
Coupling the wave property of the radiation matches

the wave property of the particle and couple to the
next higher quantum mechanical energy level

8. Remember atoms and molecules are quantum mechanical particles
9. Where a photon is a wave with some particle character, matter is made of particles
with some wave character wave/particle duality
10. As a result of this, the energy of these particles can only exist at discrete energies
we say these energy levels are quantized
11. It is easy to understand if we visualize the

wave property of matter as an oscillating
string in a boxonly certain energy levels
Energy
can exist as the string is bound at both ends:

The Spectroscopic Process
2. Absorption: Molecule takes on

the quantum energy of a photon
excited state
that matches the energy of a
transition and becomes excited 5. Detection: Relaxation as
hn photons are reemitted.
Spectrometers differ whether
they measure actual emission or
absorbance
Energy
rest state rest state
1. Irradiation: Molecule is
bombarded with photons of
hn hn hn various frequencies over
the range
CHEMdesired
430 NMR Spectroscopy 5
5
Types of Spectroscopy
core
nuclear electron electronic Nuclear Magnetic
excitation excitation excitation molecular molecular Resonance
(PET) (X-ray cryst.) (p to p*) vibration rotation NMR & MRI
g-rays
Visible
X-rays UV IR Microwave Radio
Frequency, n (Hz) ~1017 ~1015 ~1013 ~1010 ~105
Wavelength, l ~0.01 nm 10 nm 1000 nm 0.01 cm 100 m
Energy (kcal/mol) > 300 300-30 300-30 ~10-4 ~10-6
CHEM 430 NMR Spectroscopy 6 6

NMR Spectroscopy
NMR spectroscopy has emerged as the ultimate spectroscopic

method for organic structural analysis
Currently, the development of novel NMR methods is in its

golden age with some of the 2-D methods entering their
maturation period as routine spectroscopic methods
A typical NMR sample consists of 1-10 mg of sample, with which

a full analysis of 1H, 13C, DEPT, COSY, HMBC, HSQC and NOESY
could be done in a few hours on a high-field instrument
Important spin-offs of NMR spectroscopy include a host of

medical and security imaging equipment

Brief History of NMR
First NMR spectrum of H2O, 1946:
Bloch, F.; Hansen, W. W.; Packard,

M. Phys. Rev. 1946, 70 474-85.

First observation of chemical shift

1H spectrum of ethanol 1951 vs. 2011
Arnold, J.T., S.S. Dharmatti, and M.E.

Packard, J. Chem. Phys., 1951, 19, 507.

Fourier transform NMR by Ernst - 1966

2D NMR 1975 Jeener and Ernst

Brief History of NMR (MRI)
First magnetic resonance image 1973

Lauterbur and Mansfield
2011

First 3-D spectrum of small protein - 1985 Wthrich

Nobel Prizes for NMR

1944 Physics Rabi (Columbia)
1952 Physics Bloch (Stanford), Purcell (Harvard)
1991 Chemistry Ernst (ETH)
2002 Chemistry Wthrich (ETH)
2003 Medicine Lauterbur (University of Illinois in Urbana ) and
Mansfield (University of Nottingham)

Basis of NMR Spectroscopy
2.1 Magnetic Properties of Nuclei
The sub-atomic particles within atomic nuclei possess a spin
quantum number just like electrons
Just as when using Hunds rules to fill atomic orbitals with

electrons, nucleons must each have a unique set of quantum
numbers
The spin quantum number of a nucleus is a physical constant, I
For each nucleus, the total number of spin states allowed is given
by the equation:
2I + 1

Spin Quantum Numbers of Common Nuclei
Element 1H 2H 12C 13C 14N 16O 17O 19F 31P 35Cl
Nuclear Spin 1 0 1 0 5/2 3/2

Quantum Number
# of spin states 2 3 0 2 3 0 6 2 2 4
Observe that for atoms with no net nuclear spin, there are zero
allowed spin states
Nuclear Magnetic Resonance can only occur where there are

allowed spin states
Note that two nuclei, prevalent in organic compounds have

allowed nuclear spin states 1H and 13C, while two others do not
12C and 16O

There are three types of nuclei:
No spin: I = O
12C, 16O
Cannot be observed by NMR
Spinning sphere: I =
1H, 13C, 15N, 19F, 29Si, 31P)
Easiest to observe by NMR
Spinning ellipsoid I = 1, 3/2, 2

2H, 11B, 14N, 17O, 33S, 35Cl
Difficult to observe by NMR

A nucleus contains protons, which each bear a +1 charge
If the nucleus has a net nuclear spin, and an odd number of

protons, the rotation of the nucleus will generate a magnetic field
along the axis of rotation
m
I = + H H I = -
m
A hydrogen atom with its lone proton making up the nucleus, can
have two possible spin statesdegenerate in energy

The magnitude of m varies from atom to atom :
m = gI
is Plancks constant divided by 2p
g is the characteristic gyromagnetic ratio of the nucleus
The larger g is, the greater the magnetic moment

Nuclear Magnetic Resonance
When a large
In the absence
magnetic field
of stimulus all As B0 increases,
B0 is applied the
nuclear spin the larger DE
two spin states
sates are becomes
become non-
degenerate
degenerate

The large external magnetic field is defined as B0 in units of Tesla, T
The axis of B0 is defined as the z-direction
Splitting of spins into quantized groups is called the Zeeman effect
DE
CHEM 430 BNMR

0 +z Spectroscopy
direction 21
The force of B0 causes m to move in a circular motion about the z-
direction precession
B0 field in z-direction operates on the x component of m to create a

force in the y-direction (F = m X B0)
This occurs with an angular frequency w0 known as the Larmor

frequency (rad s-1)

A quantum mechanical particle can absorb a photon of energy
equal to DE and become promoted to a higher state
spectroscopic process
As B0 increases so does w0 (B0 w); the constant of

proportionality is g:
w0 = g B0
By equating w0 with Plancks relationship:

DE = hn0 = w0 = g B0

What does this mean for the NMR experiment (observing DE)?
Solving for the frequency of EM radiation we are observing:

For a bare hydrogen nucleus (H+), g = 267.53 (106 rad/Ts)
In a B0 of 1.41 Tesla:
DE = 60 MHz
DE corresponds to the highly weak radio region of the EM spectrum:
l > 5 meters and energies of < 0.02 calmol-1
This causes technical challenges to observing NMR

Boltzmann distribution more problems with NMR observation
DE at 60 MHz (DE = hn) is 2.39 x 10-5 kJ mol-1 (tiny) thermal energy at room
temperature (298 oK) is sufficient to populate both energy levels
DE is small, so rapid exchange is occurring between the two populations, but

there is always a net excess of protons in the lower energy state
From the Boltzmans Law we can calculate the population of each energy
state:
Nupper/Nlower = e-DE/kT = e-hn/kT
@ 298 oK the ratio is 1,000,000 / 1,000,009 !
There is an excess population of 9 nuclei in the lower energy state!

As the applied B0 increases, exchange becomes more difficult and the excess
increases:
Field, B0 Frequency Excess nuclei
T MHz
1.41 60 9
1.88 80 12
7.05 300 48
9.40 400 64
14.1 600 96
6. In each case, it is these few nuclei that allow us to observe NMR
7. When radio radiation is applied to a sample both transitions upward and

downward are stimulated if too much radiation is applied both states
completely equilibrate a state called saturation no observed NMR signal
2.2 Commonly Studied Nuclides

Remember that the greater DE the easier it is to detect NMR active

nuclei and have greater S/N ratios:

1. Spin
In general spin nuclei are the easiest to observe
Quadrapolar (I > ) nuclei are more difficult to observe

Unique mechanism for relaxation gives very short relaxation time
Heisenberg uncertainty principle dictates:

DE Dt ~
As relaxation times become very short, the uncertainty in

energies becomes large and peaks broaden greatly

2. Natural Abundance
Remember from Boltzmans Law we have only a small excess of nuclei
we can observe: 60 MHz, 1.41 T: 1,000,000 / 1,000,009 for 1H
1H is 99.985% of natural hydrogen - only 9 excess nuclei
Consider the excess population when the nuclei we are observing is 13C
1.11% of natural carbon
Spin couplings between low-abundant nuclei are also hampered:
Chance of two 1H-1H on adjacent Cs: 99.985 x 99.985 = 99.97%
Chance of two 13C-13C adjacent to one another: 1.11 x 1.11 = 0.1%

3. Natural Sensitivity - g
Remember - DE = hn0 = w0 = g B0
NMR signal a function of only B0 and g
The larger DE the greater the excess population for observation

(Boltzmann distribution)
1H is best followed by 19F in routine observations

4. Receptivity
Mathematical product of abundance and g
Good quick measure of amenable a nuclei is for observation
Your text uses 13C as a guidepost rather than 1H
Quick survey:
1H is 5680 times easier to observe than 13C
15N is 2.2% as easy to observe as 13C
19F is 4730 times easier to obersve than 13C; 31P is 2 times easier

2.3 The Chemical Shift
Observation of the NMR phenomenon would be of little use if all
protons resonated at the same frequency
In organic compounds protons are not bare nuclei, they are

surrounded by an s-orbital of containing an e- shared with an e- in
a hybridized orbital of another atom to form a covalent bond
In the presence of an external magnetic field, an induced

circulation of electrons opposite to that of a proton is observed
since the two are of opposite charges
This induced circulation generates a magnetic field in opposition to

the applied magnetic field a local diamagnetic current
Since the magnetic field felt by the proton within this electron cloud is
lowered, the resonance condition frequency is also lowered
This effect of lowering the energy of transition by a cloud of electrons is

called diamagnetic shielding or shielding - represented as s
The opposite effect if electron density is removed from the vicinity of

the proton is called deshielding
The actual field around the nucleus becomes B0(1 s); substitution in the
energy equation gives:

Since the magnetic field felt by the proton within this electron cloud is
lowered, the resonance condition frequency is also lowered
This effect of lowering the energy of transition by a cloud of electrons is

called diamagnetic shielding or shielding
The opposite effect if electron density is removed from the vicinity of

the proton is called deshielding
DE
CHEM 430 BNMR

0 +z Spectroscopy
direction 35
NMR Spectroscopy -
2-6
Introduction
In acetic acid, the CH3 protons
are in an e- rich environment
relative to the OH proton.
Shielding of the electrons

opposes B0 and therefore DE is
lower than that observed for
the OH proton. Here DE is
large as the full effect of B0 is
felt
DE CH3
DE OH
CHEM 430 BNMR

0 +z Spectroscopy
direction 36
DE
CHEM 430 NMR Spectroscopy

B0 37
The effect of electrons on a 1.41 T magnetic field is negligible, but
measurable
Compare the resonance frequencies for the protons in fluoromethane vs.

chloromethane
CH3F CH3Cl
The stronger inductive w/d of electrons by fluorine reduces the resonance

frequency by 72 Hz (not MHz) compared to an operating frequency of the
instrument at 60 MHz @ 1.41 T barely 1 part per million (ppm)
Using units of 60000072 vs. 60000000 is clunky at best
There needs to be a reference proton by which these chemical shifts

can be related - the best candidate would be a completely deshielded
proton (H+) which does not exist in the solution phase

NMR spectroscopists chose the other end of the spectrum- a proton that
was more shielded than any other known proton (at the time) those in
tetramethylsilane (TMS)
The 12 chemically identical protons in TMS were used as the standard zero
for an NMR spectrum
The resonance frequency of any proton to be studied (since all were less
shielded) would be at parts per million of the operating frequency of the
instrument greater than this zero
This allowed NMR instruments of varying field (and thus operating

frequency) strengths to use the same scale
Heres how:

In an applied field of 1.41 T, the resonance frequency for a typical proton is
60 MHz, at 2.35 T it is at 100 MHz a ratio of 5/3
Thus, for a given proton, the shift in Hz from the TMS standard should be 5/3
greater in the 100 MHz instrument compared to the 60 MHz
Since these are simple ratios, we can simply factor out the effect of field
strength by defining d, or chemical shift to be
d = (shift from TMS in Hz)

(spectrometer frequency in MHz)
or ppm of the instruments operating frequency

90 Mhz spectrum 300 MHz spectrum
90 Hz 300 Hz
In an applied field of 1.41 T, the resonance frequency for a typical proton is
60 MHz, at 2.35 T it is at 100 MHz a ratio of 5/3
Thus, for a given proton, the shift in Hz from the TMS standard should be 5/3
greater in the 100 MHz instrument compared to the 60 MHz
Since these are simple ratios, we can simply factor out the effect of field
strength by defining d, or chemical shift to be
d = (shift from TMS in Hz)

(spectrometer frequency in MHz)
or ppm of the instruments operating frequency

A detailed study of chemical shifts is the basis of Chapter 3
CH3
H Si
O
H3C CH
C H
C CH3 3
O
C
O H
S H H
O R C R
O H
O R
H C H R = H or alkyl
C
O
H C
C EWG
Ph
15 10 9 8 7 6 5 4 3 2 1 0.0
downfield d (ppm) upfield

deshielded shielded
higher DE lower DE

Spectrometer Design
Continuous-Wave (CW) Instrument
An NMR spectrometer needs to perform several functions:
Generate a high (>1 Tesla) magnetic field to split the energy levels of the
spin states enough to:
Create an excess nuclei population large enough to observe
Make the radio n that correspond to the transition be
observable
Ensure that the field is homogeneous (shimming)
Vary either the applied field or the radiofrequency (RF) to observe

different nuclei at their various energies of transition
Receive the faint signal of the relaxation of the excited nuclei to their
ground state
Process the signal into a usable spectrum vs. a reference

Continuous-Wave (CW) Instrument:
RF Detector
RF (60 MHz)
oscillator
Permanent
Magnet
Variable magnetic field 1.41 T few millionths of T

How it works (CW NMR):
The sample is placed in a 5 mm solution cell or tube (experimental aspects
we will cover shortly) in the center of a large permanent or electromagnet
A RF oscillator coil at 90 to the sample generates a radio signal at the

operating frequency of the instrument (60 MHz for a 1.41 T field)
The overall magnetic field is varied by a small electromagnet capping the

poles of the larger field magnet
Remember: DE = n = (g/2p) B0, so variations of either magnetic field or

frequency will cover the observed spectral width if the other is held
constant
As with older dispersive IR instruments, the sweep of magnetic fields is

simultaneous with the movement of the chart paper

How it works (CW NMR):
As a particular proton population comes into resonance, a second receiver
coil at 90 to the transmitter coil will pick up the change in orientation of
nuclear spin
This is recorded by the chart as a voltage response, proportional to the

size of the proton population that generated the resonance
One artifact of CW instruments is that the relaxation of the protons is

slower than the movement (sweep) of the chart paper
This causes the ringing effect a decreasing oscillation of the signal after
the spectrometer has moved past a given resonance
CW instruments operate by bringing each individual population of protons

into resonance individually.

Limitations- CW NMR:
Since the spectrum is collected once, the sample must possess enough
protons to give a suitable excess population that can be observed need a
concentrated sample
Due to the limitations of the relatively low magnetic field (CW instruments
top out at 60-90 MHz) the coupling constants for JHH are relatively large
compared to the spectral width so only simple molecules can be
observed and their structures elucidated
For nuclei of lower magnetogyric ratios, g, or natural abundance (13C most

specifically) the ratio of radio noise to signal is high

2.4 Excitation and Relaxation FT-NMR
Lets revisit the NMR experiment we considered earlier
Recall that at even a high applied B0 only a small excess of nuclei are in
the lower spin state (+ ) as per the Boltzman distribution
Ex: @ 7.04 T an excess of 50 spins per million
It is important to also note that 298 K imparts enough energy to the

system such that all spins are interchanging rapidly:

So when we are discussing an NMR sample we have quadrillions of
protons creating a net magnetization M in the z direction (B0)
The xy components are distributed randomly, and we can think of

the rest state as M = Mz
z z
M
y y Bo
x x
M precesses at the Larmor frequency. We are using the rotating

frame of reference to make the visualization easier.
Important note:
Each individual spin can only exist in either quantized state (
) separated by DE
The bulk magnetization however can exist at a continuum of states
M
y Bo

If a RF (w0 = w) is applied that creates a magnetic oscillation along
the x-axis a torque is applied to M rotating it towards y
z z
Mo B1
y y
B1
(or off-resonance) Mxy
wo
x x
wo
With the application of energy, a small amount of the excess

population has flipped spins. Mxy < M0

If RF energy continues to be applied the excess of + nuclei
disappears sample at saturation no NMR signal can be
detected
Natural mechanisms return an excess population to equilibrium
Any process that returns the z-magnetization to equilibrium with

an excess of + spins is called spin-lattice or longitudinal
relaxation
It is usually a first-order process and with a time constant of T1

Other magnetic nuclei in the NMR sample tumble, generating a
field from its motion
If the field is at w, excess spin energy can pass to this motional

energy as nuclei become + nuclei
For this process to be effective the nuclei need to be spatially

proximate to the tumbling molecule.
Most H-atoms are on the outside of molecules and have roughly

equal probability to relax by this process - #1Hs ~ signal

C-atoms differ substantially in their contact with external
molecules T1 relaxation usually by attached protons
Therefore T1 relaxation is more efficient for a 13CH3 rather than a

quaternary carbon or 13C=O.

T1 relaxation is responsible for generating the excess of + nuclei
in the first place!
When the sample is first placed in the B0 field, all spins are
degenerate
Magnetization builds up as spins flip from the effect of interactions

with surrounding magnetized nuclei

Any process that returns xy magnetization to its equilibrium position of
zero is called spin-spin or transverse relaxation
It is a first order process with time constant T2
By definition T2 < T1
For T2 relaxation the phases of nuclear spins must become randomized
The mechanism for this is when spin is transferred between nuclei of

opposite spin

When one spin goes from +1/2 to -1/2 while the other goes from -1/2 to
+1/2 there is no net change in z magnetization
However switches in spin cause dephasing; as the process continues xy

magnetization disappears

Detection as xy magnetization dephases the vectors begin to rotate
about z in the rotating frame of reference
A second coil 90o to the transmitter coil detects the decay in the xy
Mxy wo
x
Receiver coil (x) NMR signal

The NMR process:

Introduction to Pulse Sequences
Shorthand:
90x
Detection
Mixing time
Pulse

Pulsed Fourier Transform (FT) Instrument
First, what is a Fourier Transform?
Fourier transforms interconvert mathematical functions in the frequency domain to the

time domain:

f(n) = f(t) e-int dt
-

f(t) = p f(n) eint dt
-
For purposes of this discussion, we will black box the actual calculations and derivations
of these functions, but we need to understand what they do

Spectrometer Design
If we feed two simple oscillating equations into a FT, here are the results:
f (t) = cos (nt)
FT
-n +n
f (t) = sin (nt)
FT
+n
-n
Spectrometer Design NMR Spectroscopy

In the FT instrument, all proton populations are excited simultaneously by a short,
intense burst of RF energy
Due to a variation of the Heisenberg Uncertainty Principle, even if the RF generator

is set at 90 MHz, if the duration of the pulse is short, the radio waves do not have
time to establish a solid fundamental frequency
This can be illustrated by the following cartoon, showing the combination of a short
pulse being added to a step function:
on on
+ =
off off off off
tp = pulse duration

Spectrometer Design

4. If this short pulse is converted into the frequency domain by a FT:
on
FT
off off
tp
n
Observe that we have a continuum of frequency content centered at the
operating frequency of the instrument
We will talk more about the effects of pulse time and width when we
discuss advanced 1-D and 2-D NMR

Spectrometer Design

5. For now, if a sample containing one unique population of hydrogens was excited
over tp by a pulse, it would then relax back to its original spin state
6. As each nuclei relaxes it will emit RF radiation of a given frequency; since different
nuclei will relax at different rates, the signal decays over time
7. This emission is recorded by the spectrometer as a free-induction decay or FID
off off

Spectrometer Design

8. The actual frequency of the FID is the interference signal of the relaxing protons
superimposed with the frequency of the RF source
9. Conversion of this decay signal by FT back into the frequency domain gives us the
actual n of resonance for the proton being observed
Proton signal Pulse n

FT
time n
10.Again to due Heisenburg and other factors, NMR signals are not single lines, but a
Lorentzian shaped continuum of lines centered at the n of the signal

Spectrometer Design

Advantages:
Since all nuclei are excited and observed simultaneously, the pulse can be repeated
after each relaxation period (for 1H, about 10 seconds) and the resulting signals
added together
Because we are observing weak radiofrequency signals in a sea of RF noise for dilute
samples (or those observed once as in CW NMR) noise becomes an issue
If several to hundreds of FIDs are added together, signals will tend to constructively
add together and become more pronounced; since noise is random, it will tend to
destructively add and become less pronounced
Signal to noise ratio improves as a function of the square root of the scans (FIDs)
performed: S/N = f (n)

Back to Pulse Sequences
Standard 1-D observation of 1H or other nuclei
90x
Detection Delay
Pulse
n = scans
Delay must be longer than T1 (and T2 < T1)
Scans are repeated until S/N ratio is high enough

How can you know what T1 is?
We cannot measure it directly in the z-axis
Pulse sequence is used called inversion recovery:
180y (or x) 90y
tD

NMR
Spectroscopy 2-6 Basis of NMR Spectroscopy
tD = 0 z z
x x FT
y y
tD > 0 z z
x x FT
y y
tD >> 0 z z
x x FT
y y
NMR
If we plot the intensity versus time we get the following curve:
Intensity, (I)
time
I(t) = I * ( 1 - 2 * e - t / T1 )
It is an exponential with a time constant equal to the T1 relaxation

time.
Most 1Hs decay in less than 10 seconds no need to run routinely

NMR
How can you know what T2 is?
We cannot measure it directly like T1
Pulse sequence is used called spin-echo sequence
90y 180y (or x)
tD tD

NMR
What it does:
z y y
x tD
x
x
y
y y dephasing
tD
x
x 180y (or x)
refocusing

NMR
If we acquire the FID right after the spin-echo sequence, the intensity of
the signal after FT will only be affected by T2 relaxation and not by
dephasing due to B0 imperfections.
Upon repetition for different tD values, we plot the intensity versus 2 * tD

and get a graph similar to the one we got for inversion recovery, but in this
case the decay rate will be equal to T2.

NMR
Spectroscopy 2-6 The Coupling Constant
Consider the spectrum of ethyl alcohol:
Why does each resonance split into smaller peaks?
HO CH3
C
H2

NMR
The magnetic effects of nuclei in close proximity to those being observed
have an effect on the local magnetic field, and therefore DE
Two examples of close proximity are geminal and vicinal protons

(homonuclear) and protons attached to 13C (heteronuclear):
On any one of the 108 of these molecules in a typical NMR sample, there is
an equal statistical probability each proton is either spin + or

NMR
This creates two different magnetic environments for a proton being
observed one where its neighbor is + the other where its neighbor is

If there is more than one proton on an adjacent carbon all the statistical
probabilities exist that each one is either + or in spin
The summation of these effects over all of the observed nuclei in the
sample is observed as the spin-spin splitting of resonances

NMR
Consider 1-chloro-4-nitrobenzene
If we are observing the resonance for the HA proton,

in half the molecules in the sample HX would be
in the other half the spin would be +
We would see the resonance of HA split into two

different resonances (energy states):

NMR
Likewise, if we are observing the resonance for the

HX proton, in half the molecules in the sample HA
would be in the other half the spin would be +
We would see the resonance of HX split into two

different resonances (energy states):

NMR
The observed 1H NMR spectrum shows two doublets,

one for HA the other for HX
HA and HX are described as a spin-system

NMR Spectroscopy 2-6 The Coupling Constant
The influence of neighboring spins on peak multiplicity is called spin-spin
coupling, indirect coupling or J-coupling
The difference between the component peaks of a resonance is a measure

of how strong the interaction is between adjacent nuclei
This difference is called the coupling constant, J, measured in Hz
# of bonds
separating nuclei 3J
H-cis Nuclei that are coupled
and stereochemistry where
appropriate

NMR Spectroscopy -
Introduction 2-6 The Coupling Constant
60 MHz propyl bromide
Since J is a measure of J-values are =
interaction between two

nuclei, it must be the same for
both nuclei
J-values are =
Similarly, J is independent of
the applied B0 and will have
the same value regardless of
the field strength of the NMR

NMR Spectroscopy -
Since J is a measure of J-values are =
interaction between two

nuclei, it must be the same for
both nuclei
J-values are =
Similarly, J is independent of
the applied B0 and will have
the same value regardless of
the field strength of the NMR

NMR
Recall, we are observing the frequency (E = hn) where a proton goes into
resonance
Any change in B0 will cause a change in

energy at which the resonance condition will
occur for a proton of a given chemical shift

NMR
In solution we are not looking at a single molecule but about 108
On some molecules the proton being observed may be next to another

proton of spin + 1/2 :

NMR
proton of spin 1/2 :

NMR Spectroscopy -
Mechanism for coupling: usually oversimplified in early studies of
NMR - If molecules can rotate in solution any spatial effect of one
protons magnetic field on another is averaged to zero
The most common mechanism involves the interaction of electrons

along the bonding path between the nuclei.
Electrons, like protons, act like spinning particles and have a

magnetic moment. The X proton ( HX) influences, or polarizes, the
spins of its surrounding electrons, making the electron spins favor
one Iz state very slightly.
CHEM 430 88
NMR Spectroscopy -
Thus, a proton of spin + polarizes the electron to . The electron in
turn polarizes the other electron of the bond, and so on, finally
reaching the resonating A proton (HA).
Because J normally represents an interaction through bonds, it is a

useful parameter for drawing conclusions about molecular bonding,
such as bond order and stereochemistry.
CHEM 430 89
NMR
Observe what effect this has on an isolated ethyl group:
The two methylene Ha protons have three neighbors, Hb, on the adjacent
methyl carbon
Each one of these hydrogens can be + or , and since we are not

looking at one molecule, but billions, we will observe all combinations

NMR
The first possibility is that all three Hb protons have a + spin; in this case the
three protons combine to generate three small magnetic fields that aid B0 and
deshield the protons pushing part of the resonance for Ha slightly downfield (the
magnetic field of a proton is tiny compared to B0)
All 3 Hb
protons +
+++
Resonance, n, in absence of coupling

NMR
The second possibility is that two Hb protons have a + spin and the third a - ;
in this case the two protons combine to enhance B0 and the other against it, a net
deshielding; there are 3 different combinations that generate this state
or or
3 combinations of
+, + , - -++
+-+
+++ ++-

NMR
The third possibility is that two Hb protons have a spin and the third +; here,
the two protons combine to reduce B0 and the other enforce it, a net shielding
effect; there are 3 different combinations that generate this state
or or
3 combinations of
+, - , - -++ --+
+-+ -+-
+++ ++- +--

NMR
The last possibility is that all three Hb protons have a spin; in this case the three
protons combine to oppose B0, a net shielding effect; there is one combination
that generates this state:
All 3 Hb
protons - -++ --+
+-+ -+-
+++ ++- +-- ---

NMR
The result is instead of one resonance (peak) for Ha, the peak is split into four, a
quartet, with the constituent peaks having a ratio of 1:3:3:1 centered at the d (n)
for the resonance :
-CHa2- is split
into a quartet by
three adj. Hbs
-++ --+
+-+ -+-
+++ ++- +-- ---

NMR
Similarly, the Hb protons having two protons, on the adjacent carbon each
producing a magnetic field, cause the Hb resonance to be split into a triplet
-CHb3- is split
into a triplet by two
adj. Has
-+
++ +- --

NMR
Rather than having to do this exercise for every situation, it is quickly
recognized that a given family of equivalent protons (in the absence of
other spin-coupling) will have its resonance split into a multiplet
containing n+1 peaks, where n is the number of hydrogens on carbons
adjacent to the carbon bearing the proton giving the resonance this is
the n + 1 rule
# of Hs on Multiplet # of The relative ratios of the peaks are a mathematical
adj. Cs peaks progression given by Pascals triangle:
0 singlet 1 1
1 doublet 2 1 1
2 triplet 3 1 2 1
3 quartet 4 1 3 3 1
4 quintet 5 1 4 6 4 1
5 sextet 6 1 5 10 10 5 1
6 septet 7 1 6 15 20 15 6 1

NMR
Common patterns:
methyl - singlet tert-butyl - singlet

ethyl quartet - triplet
n-propyl triplet - quintet - triplet
iso-propyl septet - doublet

NMR

NMR

NMR
Heteronuclear coupling between 1H and 13C are not apparent in 1H spectra
(13C low abundance - 1.1%), In 99 of 100 cases, 1H are attached to
nonmagnetic 12C atoms.
In the 13C spectrum, carbon nuclei are coupled to 1H directly (99%

abundant) attached to the carbon.
Thus, the 13C resonance of a methyl carbon is split into a quartet, that of a
methylene carbon CH2 into a triplet, and that of a methine carbon CH into a
doublet. A quaternary carbon is not split by one bond coupling.
CHEM 430 101

NMR
Shown here (top) the 13C spectrum
of 3- hydroxybutyric acid which
contains a carbon resonance with
each type of multiplicity.
undecoupled
From right to left are seen a quartet
CH3 , a triplet CH2 , a doublet CH ,
and a singlet CO2H (C=O)
Instrumental procedures, called

decoupling, are available by which
spin spin split-tings may be decoupled
removed. These methods, discussed
in Section 5- 3, involve irradiating
one
CHEM 430 102

NMR
Instrumental procedures, called
decoupling, are available by which
spin spin splittings are removed for
clarity.
undecoupled
These methods involve irradiating
one nucleus with an additional field
B2 while observing another nucleus
resonating in the B1 field.
13C spectra are usually run decoupled

as other spectral techniques are decoupled
used to establish 1H-13C connectivity
more rapidly and with more clarity
CHEM 430 103

The NMR Spectrum - 1H NMR Spectroscopy
Spin-spin splitting 1H NMR
The next level of complexity (which we will cover in detail in Chapter 4) is when protons on
adjacent carbons exert different Js than one another.
Consider the ethylene fragment:
The influence of the geminal-relationship

is over the shortest distance
The magnetic influence of the trans-

relationship is over the longest distance
The cis-relationship, is over an

intermediate distance
For ethylene we would then observe three chemically distinct resonances with spin-spin
splitting exerted by the other two protons:
The observed
J couplings: multiplet for Ha
is a doublet of
2J = 0 1 Hz
gem doublets
3J
AC
3J = 11- 18 Hz
trans
3J 3J
AB AB
3J = 6 - 15 Hz
cis

Similar behavior is observed with aromatic rings; since the ring structure is fairly rigid and
electronic effects are conducted over a longer distance, J couplings are observed across
the ring system:
In low-field 1H NMR the signal for this proton

3J
would be split into a doublet by the proton
ortho ortho to it.
On a high field instrument one finds this

3J 4 5
4J
meta ortho as well as a Jmeta and a J para from
the effect of the protons meta and para to it
5J
para
Typically:
3J
ortho = 7-10 Hz
4J
meta = 1-3 Hz
5J
para = 0-1 Hz

For our initial treatment of 1H NMR the alkenyl, aromatic and the following J values should
be learned:
3J = 6-8 3J = 11-18 3J = 8-11

trans 3J = 4-8
trans
3J = 6-12
cis
3J = 6-15
cis
3J = 8-14 3J = 5-7
a,a
3J
a,e = 0-7
3J
e,e = 0-5
3J
ortho =
7-10 Hz
4J
meta = 1-3 Hz
3J = 4-10
allyl 3J = 4-8
5J
para = 0-1 Hz
trans
3J = 6-12
cis
The NMR Spectrum - 1H
A typical 1H NMR is recorded from -2 to 15 d (ppm); what is typically reported is the

region from 0 to 10 d
Remember, if a proton is shielded (e- circulation reduces felt magnetic field) DE for the
transition is lowered and the signal is near the high field or upfield region of the
spectrum (right)
If the proton is deshielded (e- circulation doesnt reduce the felt magnetic field) DE
for the transition is raised and the signal is near the low field or downfield region of
the spectrum (left)
high DE low DE
deshielded 1H sees full B0 shielded 1H reduces B0
downfield upfield
10 CHEM 430 dNMR

or ppm 0
Spectroscopy 108
The number of signals observed will be equal to the number of unique populations of
chemically equivalent protons
To determine if two protons are chemically equivalent, substitute X for that each
respective hydrogen in the compound and compare the structures
If the two structures are fully superimposible (identical) the two hydrogens are
chemically equivalent; if the two structures are different the two hydrogens were not
chemically equivalent
CH3
H
A simple example: p-xylene
CH3 X
H CH3
Same structure
H
CH3 CH3
X
H
CHEM 430 NMR Spectroscopy
CH3 109
The position (v) of each resonance is dependant on the electronic environment around
the proton chemical shift as a result of local diamagnetic shielding
There are three principle effects that contribute to local diamagnetic shielding:
1) Electronegativity
2) Hybridization
3) Proton acidity/exchange

Local Diamagnetic Shielding - Electronegativity
1. Electronegative groups comprise most organic functionalities:
-F -Cl -Br -I -OH -OR -NH2
-NHR -NR2 -NH3+ -C=O -NO2-NO -SO3H
-PO3H2 -SH -Ph -C=C and most others
In all cases, the inductive w/d of electrons of these groups decreases the electron
density in the C-H covalent bond proton is deshielded higher DE of transition

2. Protons bound to carbons bearing electron withdrawing groups are deshielded based
on the magnitude of the withdrawing effect Pauling electronegativity:
CH3F CH3O- CH3Cl CH3Br CH3I CH4 (CH3)4Si
Pauling 4.0 3.5 3.1 2.8 2.5 2.1 1.8

Electronegativity
d of H 4.26 3.40 3.05 2.68 2.16 0.23 0.0

3. The magnitude of the withdrawing effect is cumulative:
CH3Cl CH2Cl2 CHCl3

d of H 3.05 5.30 7.27
4. The magnitude of the withdrawing effect is reduced by distance, as the inductive

model suggests
-CH2Br -CH2CH2Br -CH2CH2CH2Br

d of H 3.30 1.69 1.25

Local Diamagnetic Shielding - Hybridization
1. The hybridization of the carbon the proton is bound exerts a strong electronic effect
2. The greater the s-character, the more tightly bound the electrons are to carbon, raising
its effective electronegativity (sp = 50% s, sp2, 33% s and sp3 25% s)
Type of H Name of H Chemical Shift, d
R-CH3, R2CH2, R3CH alkyl 0.8-1.7
C=C-CH3 allyl 1.6-2.6
CC-H Acetylenic 2.0-3.0
C=C-H Vinylic 4.6-5.7 Something odd

is happening
Ar-H aromatic 6.5-8.5 here, as we will
discuss
O=C-H aldehydic 9.5-10.1
Local Diamagnetic Shielding - Proton Acidity/Exchange
1. If an organic molecule that possesses hydrogen atoms of low pKA are dissolved in a
deuterated solvent that also has a low pKA, the visible protons will exchange with
deuterium from solvent and become invisible to the NMR spectrometer
OH OD
D2O
Such studies are useful, if it is desired to see which H-atoms on an organic are acidic!

Local Diamagnetic Shielding - Proton Acidity/Exchange
2. Due to H-bonding effects, the resonance for certain functional groups (esp. OH and
NH2) can change drastically dependent on concentration and the extent of the H-
bonding
3. Just as in IR spectroscopy, peaks corresponding to these resonances are broad and

often undefined observing a continuum of bond strengths/electron densities about
the observed proton
4. The correlation tables for the position of such protons tend to be broad and unreliable:
Acid OH 10.5-12.0 d
Phenol OH 4.0-12.0 d
Alcohol OH 0.5-5.0 d
Amine NH2 0.5-5.0 d
Amide NH2 5.0-8.0 d
Enol CH=CH-OH >15 d
NMR Spectroscopy
Some observed 1H resonances can not be fully explained by local diamagnetic shielding
effects
Magnetic Anisotropy literally magnetic dissimilarity
For example, by our hybridization model, a proton bound to an sp2 C should be observed
at lower d than a proton bound to an sp C
Type of H Name of H Chemical Shift, d
R-CH3, R2CH2, R3CH alkyl 0.8-1.7
C=C-CH3 allyl 1.6-2.6
CC-H Acetylenic 2.0-3.0
C=C-H Vinylic 4.6-5.7
Ar-H aromatic 6.5-8.5
O=C-H aldehydic 9.5-10.1

Magnetic Anisotropy
1. This effect is primary due to the fact that there is an additional effect of circulating
electrons, observed in p-systems
2. In benzene, the 6-p-orbitals overlap to allow full circulation of electrons; as these

electrons circulate in the applied magnetic field they oppose the applied magnetic field
at the center just like the circulation of electrons in the 1-s orbital about hydrogen
at the middle!:
B0

Magnetic Anisotropy
3. On the periphery of the ring, the effect is opposite the magnetic effect reinforces the
applied B0, and DE becomes greater deshielding effect

Magnetic Anisotropy
4. This theory can easily be tested by the observation of large aromatic systems that
possess protons inside the ring (now a shielding effect):
-1.8 d
H
8.9 d H
Or over a ring system: 2.0 d

H2
C
CH2 -1.0 d
Magnetic Anisotropy
5. In alkynes, a similar situation (to the central protons in large aromatic systems) arises
where the terminal proton is in the region of maximum shielding

General Correlation Chart 1H NMR
Due to the three effects on local diamagnetic shielding, in conjunction with the effect of
magnetic anisotropy 1H NMR chemical shifts are variable
Avoid using hard and fast rules (tables of numbers)
Instead, start from the general correlation table and deduce structural features
based on the effects just discussed
After a structural inference has been made, then use the more specific correlation
tables to confirm the analysis

General Correlation Chart 1H NMR
Here are the general regions for 1H chemical shifts: CH3
H Si
O
H3C CH
C H
C CH3 3
O
C
O H
S H H
O R C R
O H
O R
H C H R = H or alkyl
C
O
H C C EWG
Ph
15 10 9 8 7 6 5 4 3 2 1 0.0
downfield d (ppm) upfield

deshielded shielded
higher DE lower DE
1. The magnetic effects of nuclei in close proximity to those being observed have an
effect on the local magnetic field, and therefore DE
2. Specifically, when proton is close enough to another proton, typically by being on an

adjacent carbon (vicinal), it can feel the magnetic effects generated by that proton
3. On any one of the 108 of these molecules in a typical NMR sample, there is an equal
statistical probability that the adjacent (vicinal) proton is either in the + or spin
state
4. If there is more than one proton on an adjacent carbon all the statistical probabilities
exist that each one is either + or in spin
5. The summation of these effects over all of the observed nuclei in the sample is
observed as the spin-spin splitting of resonances

Intensity of SignalsIntegration
The area under an NMR signal is proportional to the number of absorbing

protons
An NMR spectrometer automatically integrates the area under the peaks, and
prints out a stepped curve (integral) on the spectrum
The height of each step is proportional to the area under the peak, which in
turn is proportional to the number of absorbing protons
Modern NMR spectrometers automatically calculate and plot the value of

each integral in arbitrary units
The ratio of integrals to one another gives the ratio of absorbing protons in a
spectrum; note that this gives a ratio, and not the absolute number, of
absorbing protons
Integration 1H NMR
1. Like instrumental chromatography, in NMR spectroscopy, the area under a peak (or
multiplet) is proportional to the number of protons in the sample that generated that
particular resonance
2. The NMR spectrometer typically will print this information on the spectrum as an
integral line (stepped line on the spectrum below)
3. The height of the integral is proportional to that proton population; by comparing the
ratios of the integrals on an NMR spectrum you can determine the number of protons
as a least common multiple of these ratios

Integration 1H NMR
4. For example observe the integration of the ethanol spectrum below:
HO CH3
C -CH3
H2
3.75 units high
-OH
1.25 units high
2.5 units high



When a nuclei of
spin + The nuclei tips The nuclei
encounters a its spin state and relaxes and
photon where is now opposed returns to the
n = E/h, the two to B0 + spin state
couple

2.1 Excellent Simulators for NMR Phenomenon
2-D compass needle analogy:
http://www.drcmr.dk/JavaCompass/
3-D Bloch equation simulator
http://www.drcmr.dk/BlochSimulator/

General Theory NMR Spectroscopy
Nuclear Spin States

The sub-atomic particles within atomic nuclei possess a spin quantum number just
like electrons
As with electrons, the nucleons are organized in energy levels
Just as when using Hunds rules to fill atomic orbitals with electrons, nucleons must
each have a unique set of quantum numbers
The total spin quantum number of a nucleus is a physical constant, I
For each nucleus, the total number of spin states allowed is given by the equation:
2I + 1

Nuclear Spin States
Spin Quantum Numbers of Common Nuclei
Element 1H 2H 12C 13C 14N 16O 17O 19F 31P 35Cl
Nuclear Spin 1 0 1 0 5/2 3/2

Quantum Number
# of spin states 2 3 0 2 3 0 6 2 2 4
6. Observe that for atoms with no net nuclear spin, there are zero allowed spin states
7. All the spin states of a given nucleus are degenerate in energy

Nuclear Magnetic Moments

A nucleus contains protons, which each bear a +1 charge
If the nucleus has a net nuclear spin, and an odd number of protons, the rotation of
the nucleus will generate a magnetic field along the axis of rotation
Thus, a nucleus has a magnetic moment, m, generated by its charge and spin
A hydrogen atom with its lone proton making up the nucleus, can have two possible
spin states, degenerate in energy
m m
+ H H -

General Theory
Nuclear Spin States

5. In the presence of an externally applied magnetic field, these two spin states are no
longer degenerate in energy
6. The spin opposed orientation is slightly higher in energy than the spin aligned
orientation
m
H -
DE B0 externally applied magnetic field
m
+ H
General Theory
Absorption of Energy
The energy difference between the two non-degenerate spin states in the presence
of an applied magnetic field is quantized
At low B0 is easy to surmise that the potential energy of the spin opposed state
would be low, and as B0 grows in strength, so would the potential energy
Thus, with increasing strength of B0, DE between the two spin states also increases
-1/2
-1/2 +1/2
DE
+1/2
CHEM 430 BNMR

0 increasing
Spectroscopy 136
4. From theory we have already discussed, we say that a quantum mechanical particle
can absorb a photon of energy equal to DE and become promoted to the higher
state
5. This energy is proportional to the frequency of the photon absorbed, and in the
case of nuclear spin, is a function of the magnetic field applied:
DE = hn = f (B0)
6. Every nucleus has a different ratio of m to angular momentum (each has a different
charge and mass) this is referred to as the magnetogyric ratio, g
DE = hn = f (gB0)
Angular momentum is quantized in units of h/2p, thus:
DE = hn = g (h/2p)B
CHEM
0 430 NMR Spectroscopy 137
General Theory
7. Solving for the frequency of EM radiation we are observing:
DE = n = (g/2p) B0
8. For a bare hydrogen nucleus (H+), g = 267.53 (106 radians/Tsec)
9. In a field strength of 1 Tesla, DE = 42.5 MHz (for our discussion, at 1.41 T, DE = 60

MHz)
DE = hn = f (B0)
This energy difference corresponds to the highly weak radio frequency region of the
EM spectrum with wavelengths of >5 meters equal to < 0.02 calmol-1
g-rays X-rays UV IR Microwave Radio

General Theory
Mechanism of absorption nuclear magnetic resonance

What we are actually observing for DE is the precessional or Larmor frequency (w)
of the spinning nucleus this is analogous to a spinning toy top precessing as a
result of the influence of the earths magnetic field:
B0

Mechanism of absorption Nuclear Magnetic Resonance

2. When a photon of n = 60 MHz encounters this spinning charged system (a bare
proton) the two can couple and change the spin state of the proton
This state is called nuclear DE w

magnetic resonance, and the
nucleus is said to be in
resonance with the incoming
radio wave
B0
n=w
w


3. The energy difference corresponding to 60 MHz (DE = hn) is 2.39 x 10-5 kJ mol-1
(tiny) thermal energy at room temperature (298 oK) is sufficient to populate both
energy levels
4. The energy difference is small, so rapid exchange is occurring between the two
populations, but there is always a net excess of protons in the lower energy state
5. From the Boltzman distribution equation we can calculate the population of each
energy state:
Nupper/Nlower = e-DE/kT = e-hn/kT
@ 298 oK the ratio is 1,000,000 / 1,000,009 !
There is an excess population of 9 nuclei in the lower energy state!

General Theory

6. As the applied B0 increases, exchange becomes more difficult and the excess
increases:
Frequency (MHz) Excess nuclei
60 9
80 12
100 16
200 32
300 48
600 96
7. In each case, it is these few nuclei that allow us to observe NMR
8. When radio radiation is applied to a sample both transitions upward and downward
are stimulated if too much radiation is applied both states completely equilibrate
a state called saturation no NMR signal can be observed

Chemical Shift
6. Spectroscopic observation of the NMR phenomenon would be of little use if all
protons resonated at the same frequency
7. The protons in organic compounds are not bare nuclei, they are surrounded by an s
-orbital of containing an electron shared with an electron in a hybridized orbital of
another atom to form a covalent bond
8. In the presence of an external magnetic field, an induced circulation of electrons

opposite to that of a proton is observed since the two are of opposite charges
9. This induced circulation generates a magnetic field in opposition to the applied

magnetic field a local diamagnetic current


The magnitude of m from atom to atom varies:
m = gI
is Plancks constant divided by 2p
g is the characteristic gyromagnetic ratio of the nucleus
The larger g is, the greater the magnetic moment

For the 1H nucleus (proton) this resonance condition occurs at low

energy (lots of noise) unless a very large magnetic field is applied
Early NMR spectrometers used a large permanent magnet with a

field of 1.4 Teslaprotons undergo resonance at 60 MHz (1 MHz =
106 Hz)
Modern instruments use a large superconducting magnetour

NMR operates at 9.4 T where proton resonance occurs at 400 MHz
In short, higher field gives cleaner spectra and allows longer and
more detailed experiments to be performed

Origin of the Chemical Shift
Electrons Deshieding Shielding

surrounding the Factors which lower e- Factors which raise
nucleus are density allow the e- density reduce
opposite in charge nucleus to see more the amount of B0
to the proton, of the B0 being the nucleus sees
therefore they applied resonance resonance
generate an occurs at higher condition occurs at
opposing b0 energy lower energy

The Proton (1H) NMR Spectrum

The 1H NMR Spectrum
A reference compound is neededone that is inert and does not

interfere with other resonances
Chemists chose a compound with a large number of highly shielded

protonstetramethylsilane (TMS)
No matter what spectrometer is used the resonance for the protons on

this compound is set to d 0.00
CH3
Si
H3C CH
CH3 3

The 1H NMR Spectrum
The chemical shift for a given proton is in frequency units (Hz)
This value will change depending on the B0 of the particular spectrometer
By reporting the NMR absorption as a fraction of the NMR operating

frequency, we get units, ppm, that are independent of the spectrometer

The 1H NMR Spectrum
We need to consider four aspects of a 1H spectrum:
a. Number of signals
b. Position of signals
c. Intensity of signals.
d. Spin-spin splitting of signals

The Number of Signals
The number of NMR signals equals the number of different types of

protons in a compound
Protons in different environments give different NMR signals
Equivalent protons give the same NMR signal

To determine if two protons are chemically equivalent, substitute X for

that each respective hydrogen in the compound and compare the
structures
If the two structures are fully superimposible (identical) the two

hydrogens are chemically equivalent; if the two structures are different
the two hydrogens were not equivalent
CH 3
A simple example: p-xylene H
CH 3 Z
H CH 3
Same Compound
H
CH 3 CH 3
Z
CHEM 430 NMR Spectroscopy H

152
CH 3
Examples
Important: To determine equivalent protons in cycloalkanes and alkenes, always draw

all bonds to show specific stereochemistry:

In comparing two H atoms on a ring or double bond, two protons are

equivalent only if they are cis or trans to the same groups.

Proton equivalency in cycloalkanes can be determined similarly:

Enantiotopic Protons when substitution of two H atoms by Z forms

enantiomers:
a. The two H atoms are equivalent and give the same NMR signal
b. These two atoms are called enantiotopic

Diastereotopic Protons - when substitution of two H atoms by Z forms

diastereomers
a. The two H atoms are not equivalent and give two NMR signals
b. These two atoms are called diastereotopic

Chemical Shift Position of Signals
Remember:
Electrons Deshieding Shielding

surrounding the Factors which lower Factors which raise
nucleus are e- density allow the e- density reduce the
opposite in charge nucleus to see amount of B0 the
to the proton, more of the B0 being nucleus sees
therefore they applied resonance resonance condition
generate an occurs at higher occurs at lower
opposing b0 energy energy
The less shielded the nucleus becomes, the more of the applied magnetic
field (B0) it feels
This deshielded nucleus experiences a higher magnetic field strength, to it

needs a higher frequency to achieve resonance
Higher frequency is to the left in an NMR spectrum, toward higher

chemical shiftso deshielding shifts an absorption downfield
Downfield, deshielded Upfield, shielded

There are three principle effects that contribute to local

diamagnetic shielding:
a. Electronegativity
b. Hybridization
c. Proton acidity/exchange

Electronegative groups comprise most organic functionalities:
-F -Cl -Br -I -OH -OR -NH2
-NHR -NR2 -NH3+ -C=O -NO2-NO -SO3H
-PO3H2 -SH -Ph -C=C and most others
In all cases, the inductive WD of electrons of these groups

decreases the electron density in the C-H covalent bond
proton is deshielded signal more downfield of TMS

Protons bound to carbons bearing electron withdrawing groups are

deshielded based on the magnitude of the withdrawing effect Pauling
electronegativity:
CH3F CH3O- CH3Cl CH3Br CH3I CH4 (CH3)4Si
Pauling 4.0 3.5 3.1 2.8 2.5 2.1 1.8

Electronegativity
d of H 4.26 3.40 3.05 2.68 2.16 0.23 0.0

3. The magnitude of the deshielding effect is cumulative:
CH3Cl CH2Cl2 CHCl3

d of H 3.05 5.30 7.27
As more chlorines are added d becomes larger
3. The magnitude of the deshielding effect is reduced by distance, as the

inductive model suggests
-CH2Br -CH2CH2Br -CH2CH2CH2Br

d of H 3.30 1.69 1.25

Hybridization
Increasing s-character (sp3 sp2 sp) pulls e- density closer to nucleus
effectively raising electronegativity of the carbon the H atoms are bound
to a deshielding effect
We would assume that H atoms on sp carbons should be well downfield

(high d) and those on sp3 carbons should be upfield (low d)

What we observe is slightly different:
Type of H Carbon Name of H Chemical Shift, d

hybridization
R-CH3, R2CH2, R3CH sp3 alkyl 0.8-1.7
C=C-CH3 sp3 allyl 1.6-2.6
CC-H sp acetylenic 2.0-3.0
C=C-H sp2 vinylic 4.6-5.7
Ar-H sp2 aromatic 6.5-8.5
O=C-H sp2 aldehydic 9.5-10.1
Chemists refer to this observation as magnetic anisotropy

Magnetic Anisotropy Aromatic Protons

a. In a magnetic field, the six p electrons in benzene circulate around
the ring creating a ring current.
b. The magnetic field induced by these moving electrons reinforces the
applied magnetic field in the vicinity of the protons.
c. The protons thus feel a stronger magnetic field and a higher
frequency is needed for resonance. Thus they are deshielded and
absorb downfield.

Similarly this effect operates in alkenes:

In alkynes there are two perpendicular sets of p-electronsthe molecule

orients with the field lengthwiseopposing B0 shielding the terminal H atom



The area under an NMR signal is proportional to the number of

absorbing protons
An NMR spectrometer automatically integrates the area under the

peaks, and prints out a stepped curve (integral) on the spectrum
The height of each step is proportional to the area under the peak,
which in turn is proportional to the number of absorbing protons
Modern NMR spectrometers automatically calculate and plot the value

of each integral in arbitrary units
The ratio of integrals to one another gives the ratio of absorbing

protons in a spectrum; note that this gives a ratio, and not the absolute
number, of absorbing protons



Spin-Spin Splitting
Consider the spectrum of ethyl alcohol:

Why does each resonance split into smaller peaks?
HO CH3
C
H2

Spin-Spin Splitting
The magnetic effects of nuclei in close proximity to those being

observed have an effect on the local magnetic field, and therefore DE
Specifically, when proton is close enough to another proton, typically

by being on an adjacent carbon (vicinal), it can feel the magnetic
effects generated by that proton
On any one of the 108 of these molecules in a typical NMR sample,

there is an equal statistical probability that the adjacent (vicinal)
proton is either in the + or spin state
If there is more than one proton on an adjacent carbon all the

statistical probabilities exist that each one is either + or in spin
The summation of these effects over all of the observed nuclei in the
sample is observed as the spin-spin splitting of resonances

Spin-Spin Splitting
Recall, we are observing the frequency (E = hn) where a proton goes

into resonance
Any change in B0 will cause a change in

energy at which the resonance condition will
occur for a proton of a given chemical shift

In solution we are not looking at a single molecule but about 108
proton of spin + 1/2 :

Spin-Spin Splitting
On some molecules the proton being observed may be next to

another proton of spin 1/2 :

Spin-Spin Splitting
Observe what effect this has on an isolated ethyl group:
The two methylene Ha protons have three neighbors, Hb, on the

adjacent methyl carbon
Each one of these hydrogens can be + or , and since we are not

looking at one molecule, but billions, we will observe all combinations

Spin-Spin Splitting
The first possibility is that all three Hb protons have a + spin; in this
case the three protons combine to generate three small magnetic fields
that aid B0 and deshield the protons pushing the resonance for Ha
slightly downfield (the magnetic field of a proton is tiny compared to B0)
All 3 Hb
protons +

resonance for Ha in absence of spin-spin splitting
Spin-Spin Splitting
The second possibility is that two Hb protons have a + spin and the third
a - ; in this case the two protons combine to enhance B0 and the other
against it, a net deshielding;
there are 3 different combinations that generate this state
or or
2 Hb
protons +

Spin-Spin Splitting
The third possibility is that two Hb protons have a spin and the third
+; here, the two protons combine to reduce B0 and the other enforce
it, a net shielding effect;
there are 3 different combinations that generate this state
or or
2 Hb
protons -

Spin-Spin Splitting
The last possibility is that all three Hb protons have a spin; in this case
the three protons combine to oppose B0, a net shielding effect;
there is one combination that generates this state
All 3 Hb
protons -

Spin-Spin Splitting
The result is instead of one resonance (peak) for Ha, the peak is split
into four, a quartet, with the constituent peaks having a ratio of 1:3:3:1
centered at the d (n) for the resonance

Spin-Spin Splitting
Similarly, the Hb protons having two protons, on the adjacent carbon each
producing a magnetic field, cause the Hb resonance to be split into a
triplet

Spin-Spin Splitting
Rather than having to do this exercise for every situation, it is quickly

recognized that a given family of equivalent protons (in the absence of other
spin-coupling) will have its resonance split into a multiplet containing n+1
peaks, where n is the number of hydrogens on carbons adjacent to the carbon
bearing the proton giving the resonance this is the n + 1 rule
# of Hs Multiplet # of The relative ratios of the peaks are a

on adj. peaks mathematical progression given by Pascals
Cs triangle:
0 singlet 1 1
1 doublet 2 1 1
2 triplet 3 1 2 1
3 quartet 4 1 3 3 1
4 quintet 5 1 4 6 4 1
5 sextet 6 1 5 10 10 5 1
6 septet 7 CHEM 430 NMR Spectroscopy 1 6 15 20 15 6 1 186
1H NMRSpin-Spin Splitting
Common patterns:
methyl - singlet tert-butyl - singlet

ethyl quartet - triplet
n-propyl triplet - quintet - triplet
iso-propyl septet - doublet


Another Example:
Br
Br C
Br = Br
Br C
Br Ha
Hb Hb

Another Example:

Three general rules describe the splitting patterns commonly

seen in the 1H NMR spectra of organic compounds:
1.Equivalent protons do not split each others signals
2.A set of n nonequivalent protons splits the signal of a
nearby proton into n + 1 peaks
3.Splitting is observed for nonequivalent protons on the same
carbon or adjacent carbons
If Ha and Hb are not equivalent, splitting is observed when:

Magnetic influence falls off dramatically with distance
The n + 1 rule only works in the following situations:
H H
Aliphatic compounds that have free
rotation about each bond
H H
G
H Ha
Aromatic compounds where each proton is
H Hb held in position relative to one another
Hc CHEM 430 NMR Spectroscopy 192
The amount of influence exerted by a proton on an adjacent

carbon is observed as the difference (in Hz) between component
peaks within the multiplet it generates. This influence is
quantified as the coupling constant, J
Two sets of protons that split one another are said to be coupled
J for two sets of protons that are coupled are equivalent

therefore on complex spectra we can tell what is next to what
This J Is equal to this J
-CH2- CHEM 430 NMR Spectroscopy -CH3 193

The next level of complexity (which at this level, is only introduced) is

when protons on adjacent carbons exert different Js than one another.
Consider the ethylene fragment:

The influence of the geminal-relationship is
over the shortest distance
The magnetic influence of the trans-

relationship is over the longest distance
The cis-relationship, is over an

intermediate distance
For this substituted ethylene we see the following spectrum:
The observed multiplet

for Ha is a doublet of
2J = 0 1 Hz
gem doublets
3J
AC
3J = 11- 18 Hz
trans
3J 3J
AB AB
3J = 6 - 15 Hz
cis

In general, when two sets of adjacent protons are different from each
other (n protons on one adjacent carbon and m protons on the other),
the number of peaks in an NMR signal = (n + 1)(M + 1)
In general the value of J falls off with distance; J values have been
tabulated for virtually all alkene, aromatic and aliphatic ring systems

Some common J-values
3J = 6-8 3J = 11-18 3J = 8-11

trans 3J = 4-8
trans
3J = 6-12
cis
3J = 6-15
cis
3J = 8-14 3J = 5-7
a,a
3J
a,e = 0-7
3J
e,e = 0-5
3J
ortho = 7-10 Hz
4J
meta = 1-3 Hz
3J = 4-10 5J
allyl 3J
trans= 4-8 para = 0-1 Hz
3J = 6-12
cis
We can now tell stereoisomers apart through 1H NMR:

A combined example:

Under usual conditions, an OH proton does not split the NMR signal
of adjacent protons
Protons on electronegative atoms rapidly exchange between
molecules in the presence of trace amounts of acid or base (usually
with NH and OH protons)

Structure Determination




13C NMR
The lack of splitting in a 13C spectrum is a consequence of the low

natural abundance of 13C
Recall that splitting occurs when two NMR active nucleilike two
protonsare close to each other. Because of the low natural
abundance of 13C nuclei (1.1%), the chance of two 13C nuclei being
bonded to each other is very small (0.01%), and so no carbon-
carbon splitting is observed
A 13C NMR signal can also be split by nearby protons. This 1H-13C
splitting is usually eliminated from the spectrum by using an
instrumental technique that decouples the proton-carbon
interactions, so that every peak in a 13C NMR spectrum appears as a
singlet
The two features of a 13C NMR spectrum that provide the most
structural information are the number of signals observed and the
chemical shifts of those signals

13C NMR

13C NMR
The number of signals in a 13C spectrum gives the number of different

types of carbon atoms in a molecule.
Because 13C NMR signals are not split, the number of signals equals the
number of lines in the 13C spectrum.
In contrast to the 1H NMR situation, peak intensity is not proportional to
the number of absorbing carbons, so 13C NMR signals are not integrated.

13C NMR
In contrast to the small range of chemical shifts in 1H NMR (1-10

ppm usually), 13C NMR absorptions occur over a much broader range
(0-220 ppm).
The chemical shifts of carbon atoms in 13C NMR depend on the same
effects as the chemical shifts of protons in 1H NMR.

13C NMR

13C NMR

Shoolery Tables
After years of collective observation of 1H and 13C NMR it is possible to

predict chemical shift to a fair precision using Shoolery Tables
These tables use a base value for 1H and 13C chemical shift to which are
added adjustment increments for each group on the carbon atom
H H H
X C H X C Y X C Z
H H Y
methyl methylene methine

Shoolery Values for Methylene
X or Y Substituent X or Y Substituent
Constant Constant
-H 0.34 -OC(=O)OR 3.01
-CH3 0.68 -OC(=O)Ph 3.27
-CC 1.32 -C(=O)R 1.50
-CC- 1.44 -C(=O)Ph 1.90
-Ph 1.83 -C(=O)OR 1.46
-CF2- 1.12 -C(=O)NR2 or 1.47
H2
-CF3 1.14 -CN 1.59
-F 3.30 -NR2 or H2 1.57
-Cl 2.53 -NHPh 2.04
-Br 2.33 -NHC(=O)R 2.27
-I 2.19 CHEM 430 NMR Spectroscopy
-N 1.97 212
3
-OH 2.56 -NO2 3.36

Shoolery Values for Methine
X ,Y or Z Substituent X, Y or Z Substituent
Constant Constant
-F 1.59 -OC(=O)OR 0.47
-Cl 1.56 -C(=O)R 0.47
-Br 1.53 -C(=O)Ph 1.22
-NO2 1.84 -CN 0.66
-NR2 or H2 0.64 -C(=O)NH2 0.60
-NH3+ 1.34 -SR or H 0.61
- 1.80 -OSO2R 0.94
NHC(=O)R
-OH 1.14 -CC- 0.79
-OR 1.14 -C=C 0.46
-C(=O)OR 2.07 -Ph 0.99

-OPh 1.79
Shoolery Tables
For methyluse methylene formula and table

using the H value
For methyleneuse a base value of 0.23 and

add the two substituent constants for X and Y
In 92% of cases experimental is within 0.2 ppm
For methineuse a base value of 2.50 and add

the three substituent constants for X, Y and Z
Error similar to methylene
Shoolery Tables
Work for aromatics as well (.pdf posted)

Running an NMR Experiment
Sample sizes for a typical high-field NMR (300-600 MHz):

1-10 mg for 1H NMR
10-50 mg for 13C NMR
Solution phase NMR experiments are much simpler to run; solid-

phase NMR requires considerable effort
Sample is dissolved in ~1 mL of a solvent that has no 1H hydrogens
Otherwise the spectrum would be 99.5% of solvent, 0.5% sample!

Deuterated solvents are employedall 1H atoms replaced with 2H

which resonates at a different frequency
Most common: CDCl3 and D2O
Employed if necessary: CD2Cl2, DMSO-d6, toluene-d8, benzene-d6,

CD3OD, acetone-d6
Sample is contained in a high-tolerance

thin glass tube (5 mm)

IMPORTANTno deuterated solvent is 100% deuterated, there is

always residual 1H material, and this will show up on the spectrum
CHCl3 in CDCl3 is a singlet at d 7.27
HOD in D2O is a broad singlet at d 4.8
No attempt is made to make solvents for 13C NMR free of 13C, as the
resonances are so weak to begin with
13C NMR using CDCl3 shows a unique 1:1:1 triplet at d 77.00 (+1, 0, 1
spin states of deuterium coupled with 13C)

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Introduction to Spectroscopy

Spectroscopy is the study of the interaction of matter with the

1. EM radiation displays the properties of both particles and waves

2. This packet of wave and particle properties is called a photon

The term photon is implied to mean a small, massless particle that

3. The energy E component of a photon is proportional to the frequency

The constant of proportionality is Planks constant, h

CHEM 430 NMR Spectroscopy 2

4. Because the speed of light (c ) is constant, the frequency (n)

6. Because the atomic particles in matter also exhibit wave and

Coupling the wave property of the radiation matches

CHEM 430 NMR Spectroscopy 3

8. Remember atoms and molecules are quantum mechanical particles

11. It is easy to understand if we visualize the

CHEM 430 NMR Spectroscopy 4

2. Absorption: Molecule takes on

rest state rest state

Frequency, n (Hz) ~1017 ~1015 ~1013 ~1010 ~105

Wavelength, l ~0.01 nm 10 nm 1000 nm 0.01 cm 100 m

Energy (kcal/mol) > 300 300-30 300-30 ~10-4 ~10-6

CHEM 430 NMR Spectroscopy 6 6

NMR spectroscopy has emerged as the ultimate spectroscopic

Currently, the development of novel NMR methods is in its

A typical NMR sample consists of 1-10 mg of sample, with which

Important spin-offs of NMR spectroscopy include a host of

CHEM 430 NMR Spectroscopy 7

Bloch, F.; Hansen, W. W.; Packard,

CHEM 430 NMR Spectroscopy 8

First observation of chemical shift

Arnold, J.T., S.S. Dharmatti, and M.E.

CHEM 430 NMR Spectroscopy 9

Fourier transform NMR by Ernst - 1966

CHEM 430 NMR Spectroscopy 10

2D NMR 1975 Jeener and Ernst

CHEM 430 NMR Spectroscopy 11

First magnetic resonance image 1973

CHEM 430 NMR Spectroscopy 12

First 3-D spectrum of small protein - 1985 Wthrich

CHEM 430 NMR Spectroscopy 13

Nobel Prizes for NMR

CHEM 430 NMR Spectroscopy 14

Just as when using Hunds rules to fill atomic orbitals with

The spin quantum number of a nucleus is a physical constant, I

CHEM 430 NMR Spectroscopy 15

Nuclear Spin 1 0 1 0 5/2 3/2

Nuclear Magnetic Resonance can only occur where there are

Note that two nuclei, prevalent in organic compounds have

CHEM 430 NMR Spectroscopy 16

Cannot be observed by NMR

Easiest to observe by NMR

Spinning ellipsoid I = 1, 3/2, 2

Difficult to observe by NMR

CHEM 430 NMR Spectroscopy 17

If the nucleus has a net nuclear spin, and an odd number of

CHEM 430 NMR Spectroscopy 18

is Plancks constant divided by 2p

g is the characteristic gyromagnetic ratio of the nucleus

The larger g is, the greater the magnetic moment

CHEM 430 NMR Spectroscopy 19

CHEM 430 NMR Spectroscopy 20

The axis of B0 is defined as the z-direction

Splitting of spins into quantized groups is called the Zeeman effect

CHEM 430 BNMR

B0 field in z-direction operates on the x component of m to create a