Nuclear Magnetic Resonance (NMR) Spectroscopy

 Basic principles of NMR

 Chemical shift scale
 Different types of H
 Shielding in H-NMR
o due to electronegativity
o magnetic anisotropy
o hydrogen bonding
 Table of H-NMR chemical shifts
 H-NMR spectra I
 Coupling in H-NMR
 H-NMR spectra II
 Complex coupling patterns
 Interpreting H-NMR
 C-NMR spectra basics
 Interpreting C-NMR
 DEPT C-NMR
 2D-NMR

Basics
Nuclei with an odd mass or odd atomic number have "nuclear spin" (in a
similar fashion to the spin of electrons). This includes 1H and 13C
(but not 12C). The spins of nuclei are sufficiently different that NMR
experiments can be sensitive for only one particular isotope of one
particular element. The NMR behaviour of 1H and 13C nuclei has been
exploited by organic chemist since they provide valuable information that
can be used to deduce the structure of organic compounds. These will be
the focus of our attention.

Since a nucleus is a charged particle in motion, it will develop a magnetic

field. 1H and 13C have nuclear spins of 1/2 and so they behave in a similar
fashion to a simple, tiny bar magnet. In the absence of a magnetic field,
these are randomly oriented but when a field is applied they line up parallel
to the applied field, either spin aligned or spin opposed. The more highly
populated state is the lower energy spin state spin aligned situation. Two
schematic representations of these arrangements are shown below:
Basic Principles
In NMR, electromagnetic (EM) radiation is
used to "flip" the alignment of nuclear spins
from the low energy, spin aligned state to
the higher energy spin opposed state.

The energy required for this transition

depends on the strength of the applied
magnetic field (see below). The energy
difference between the spin states is quite
small and corresponds to the radio
frequency range of the EM spectrum.
As this diagram shows, the energy required to cause the
spin-flip, DE, depends on the magnetic field strength at the
nucleus. With no applied field, there is no energy difference
between the spin states, but as the field increases, so does the
separation of energies of the spin states and therefore so does
the frequency required to cause the spin-flip.

DE' > DE since Ho' > Ho

The basic arrangement

of an NMR
spectrometer is shown
to the left. The sample
is positioned in the
magnetic field and
excited via pulsations
in the radio frequency
input circuit. The
realigned magnetic
fields induce a radio
signal in the output
circuit which is used to
generate the output
signal. Fourier analysis
of the complex output
produces the actual
spectrum. The pulse is
repeated as many times
as necessary to allow
the signals to be
identified from the
background noise.

Chemical Shift
 An NMR spectrum is a plot of the radio frequency
applied against absorption.
 A signal in the spectrum is referred to as a resonance.
 The frequency of a signal is known as its chemical shift, 
The chemical shift in absolute terms is defined by the frequency of the resonance
expressed with reference to a standard compound which is defined to be at 0 ppm.
The scale is made more manageable by expressing it in parts per million (ppm) and
is indepedent of the spectrometer frequency.

It is often convienient to describe the relative positions of the resonances in an NMR

spectrum. For example, a peak at a chemical shift, δ, of 10 ppm is said to
be downfield or deshielded with respect to a peak at 5 ppm, or if you prefer, the
peak at 5 ppm is upfield or shielded with respect to the peak at 10 ppm. The
terms shielded and deshielded will be explained later.

Typically for a field strength of 4.7 T the resonance frequency of a proton

will occur around 200MHz and for a carbon, around 50.4MHz.
The reference compound is the same for both, tetramethysilane (Si(CH3)4).

Different types of Hydrogen

The issues to understand from this page are:

 What do we mean by different types of hydrogen?

 Counting types of hydrogen
 Terminology

This may seem a trivial concept, but the idea of establishing the number of
kinds of hydrogens (or other atoms such as C) is a very important
useful concept, especially in NMR spectroscopy where for example,
different types of H typically give rise to a different signal.

What do we mean?
A hydrogen atom is "different" to another hydrogen atom if it is not in an
identical environment to the other hydrogen. This could mean it's attached
to a different type of atom (e.g. CH vs OH, or sp3 CH vs sp2 CH) or due to
the number of adjacent H (e.g. CH3- vs -CH2- ) or just at a different point in
a chain (e.g. compare the H in the methylene (CH2) groups in
CH3CH2CH2OH

Counting different types of Hydrogen

There are three methods that can be used to count the number of kinds of
H (each achieve the same result). You will probably find it best to master
the first of these methods and over time you will find yourself moving
towards the 3rd method)

1. Substitution method (simplest but slowest)

The idea is that you replace each H in the molecule in turn with a "dummy"
atom (for example a -Cl) to see if you get a different product (i.e. one that
will require a different name). Each new product, indicates a different type
of H. This idea is related to the radical chlorination of alkanes where some
of each possible product is usually obtained.

For example:

Qu : If you remove one H atom from chloroethane and substitute it for

another Cl atom, how many different molecules can you make ?
Ans : Two products, 1,1-dichloroethane and 1,2-dichloroethane. Hence
there are two types of H in chloroethane

2. Verbal description
The verbal method requires that you describe the position of each H
within the molecule. If you need to use different words to describe two
H atoms, then they represent different types of H.

For example:

 an -OH is different to a -CH (based on what they are attached to)

 a -CH3 is different to a -CH2- (because the number of H at that C are
different)
 an sp3C-H is different to an spC-H.
 Other differences could be position on a chain, across a ring or
double bond etc.

3. Symmetry (most difficult but fastest)

The symmetry method is the most sophisticated but the quickest method
and requires that you look for mirror planes, rotation axes or inversion
centers that interchange H atoms. H atoms that can be interchanged are
equivalent to each other.

Shielding in H-NMR
 Structural factors cause changes in the magnetic field experienced
by the nucleus
 As we have seen, this changes the resonance frequency and hence
the chemical shift

The structural factors mean that different types of proton will occur
at different chemical shifts. This is what makes NMR so useful for
structure determination, otherwise all protons would occur at the same
frequency, limiting the information to the presence of H.

The various factors that influence the field include:

 inductive effects by electronegative groups

 magnetic anisotropy of π-systems
 hydrogen bonding

Electronegativity
The electrons around the proton create a magnetic field that opposes the
applied field. Since this reduces the field
experienced at the nucleus, the electrons are said to shield the proton. It
can be useful to think of this in terms of vectors....
 Since the field experienced by the proton
defines the energy difference between the
two spin states, the frequency and hence the
chemical shift, δ /ppm, will change
depending on the electron density around
the proton. Electronegative groups attached
to the C-H system decrease the electron
density around the protons, and there is less
shielding (i.e. deshielding) so the chemical
shift increases. This is reflected by the plot
shown in the graph to the left which is
based on the data shown below.

Compound,
CH3F CH3OH CH3Cl CH3Br CH3I CH4 (CH3)4Si
CH3X
X F O Cl Br I H Si
Electronegativity
4.0 3.5 3.1 2.8 2.5 2.1 1.8
of X
Chemical
4.26 3.4 3.05 2.68 2.16 0.23 0
shift, δ / ppm

These effects are cumulative, so the

presence of more electronegative Compound CH4 CH3Cl CH2Cl2 CHCl3
groups produce more deshielding
and therefore, larger chemical δ / ppm 0.23 3.05 5.30 7.27
shifts.
These inductive effects are not just
felt by the immediately adjacent
protons as the disruption of electron
density has an influence further
down the chain. However, the H signal -CH2-CH2-CH2Br
effect does fade rapidly as you δ / ppm 1.25 1.69 3.30
move away from the electronegative
group. As an example, look at the
chemical shifts for part of a primary
bromide

Magnetic Anisotropy
The word "anisotropic" means "non-uniform". So magnetic anisotropy
means that there is a "non-uniform magnetic field". Electrons in π systems
(e.g. aromatics, alkenes, alkynes, carbonyls etc.) interact with the applied
field which induces a magnetic field that causes the anisotropy. As a
result, the nearby protons will experience 3 fields: the applied field, the
shielding field of the valence electrons and the field due to the π system.
Depending on the position of the proton in this third field, it can be either
shielded (smaller d) or deshielded (larger d), which implies that the energy
required for, and the frequency of the absorption will change.

Hydrogen Bonding
Protons that are involved in hydrogen bonding (this usually means -OH or -
NH) are typically observed over a large range of chemical shift values. The
more hydrogen bonding there is, the more the proton is deshielded and the
higher its chemical shift will be. However, since the amount of hydrogen
bonding is susceptible to factors such as solvation, acidity, concentration
and temperature, it can often be difficult to predict.

HINT : It is often a good idea to leave assigning -OH or -NH resonances

until other assignments have been made.

Experimentally, -OH and -NH protons can be identified by carrying out a

simple D2O (deuterium oxide, also known as heavy water) exchange
experiment. These H atoms are said to be exchangeable.
  Run the regular H-NMR experiment
 Add a few drops of D2O
 Re-run H-NMR experiment
 Compare the two spectra and look for peaks that have "disappeared"

Why would a peak disappear ?

Consider the alcohol case for example: R-OH + D2O <=> R-

OD + HOD
During the hydrogen bonding, the alcohol and heavy water can "exchange"
-H and -D each other, so the alcohol becomes R-OD.
Although D is NMR active, its signals are of different energy and are not
seen in the H-NMR, hence the peak due to the -OH disappears.

Solvents:
A solvent is a liquid substance that lets other substances to get dissolved in it.
The water is a universal solvent and is least expensive and also widely. But
there are other solvents preferred based on the need like the ethanol, oils and
petroleum products etc.
They find many applications in the formulation of food, drugs, cosmetics
(lipsticks) and also in research.

Common Examples of Solvents include

1. Water
2. Ethanol
3. Methanol
4. N-propanal
5. Butanol
6. Ether
7. Dichloromethane
8. Carbon disulfide
9. glycerol
10. Acetone
11. Carbon tetrachloride
12. Cyclohexane
13. Formic acid
14. Toluene
15. Anisole
16. Pyridine
17. Acetic Acid
18. Hexane
19. Xylene
20. Trifluoroacetic acid
21. Dimethyl sulfoxide
22. benzene
23. Nitrobenzene
24. Quinoline
25. Dibutyl phthalate
26. Dimethylformamide
27. Cyclohexane
28. Anisole
29. Tetrahydrofuran
30. Petroleum ether
Solvents are chemical compounds which are physically liquids at the room
temperature. Besides these, even gases can act as solvents when required.
In science, solvents are useful for chemical analysis by
titration, chromatography, spectrometry etc.
Where as in industry these solvents are used fundamentally for the extraction,
purification and also molding of substances into different shapes.

There are different types of solvents which are routinely used

Different types of solvents
Solvents can be briefly classified based on their chemical nature and
behavior.

A. Based upon Polarity:

In general, most solvents have polarity in their internal chemistry.
This polarity is due to the concentration of opposite charges on one of the
atom or element inside a solvent molecule.

It imparts changes on the solute molecule structure such that they get
dissolved by forming ions.

When a solute is mixed in a solvent, the solvent molecules dissolve the solute
by separating apart the solute molecules using forces like hydrogen bonding,
Vanderwal forces, etc.

Examples: Sodium chloride has NaCl molecule, which breaks into Na+ and Cl-
ions when dissolved in water.
1. Polar solvents: These are solvents having a dielectric constant of more
than 15. They can dissolve salts and other ionizable solutes. Polar solvents
examples include water, alcohol. Polar solutes like the salts dissolve in polar
solvents.
2. Non-polar solvents. These solvents are nonpolar and have dielectric
constants less than 15. They cannot form inter-molecular bonds by use of
hydrogen bonding, Vanderwal forces, etc. Hence they cannot dissolve polar
compounds. Non polar solvents examples include Benzene, CCl4.
Fats and oils are soluble in non-polar solvents. Hence to remove lipids from
an extract, petroleum ether is used in the industry.
B. Based on Chemical nature:
1. Aprotic solvents: (No protons). These solvents are nonreactive and
chemically inert. They neither take protons nor give protons. Ex: benzene
(C6H6). Chloroform (CHCl3).
2. Amphiprotic solvents: Theses solvents which can provide and take up
protons on reaction. They have a neutral pH. Ex: Water, alcohol.
3. Protogenic solvents (proton+genesis = give): These are the solvents
acidic by nature. They can donate a proton and hence called as “protogenic.”
Ex: HCL, H2SO4, perchloric acid.
4. Protophyllic solvents: These are the solvents which take up protons. They
are basic by nature and are mostly alkalies. Ex: NaOH, KOH, etc.
These the and protophilic solvents can be again classified as leveling agents
and differentiating agents.
A strong acid or base is leveling agents as it can donate or accept protons to
even weak base or acid respectively.

While weak acids and weak bases cannot do so, they can only give proton to
strong base or take up a proton from strong acid respectively. Hence due to
this differentiation, they are called differentiating agents.

C) Based on chemistry:
Solvents are also classified based on their center of chemistry due to the
presence of some particular elements. These unique elements in
solvents bring a total change in their physical and chemical properties.
Inorganic solvents: Solvents without carbon are called inorganic solvents.
Ex: water, NaOH, HCl
Organic solvents. Solvents having carbon are called organic solvents. Ex:
Alcohols (CH3OH), hydrocarbons solvents like Benzene.
Halogenated solvents: Solvents having halogens are called halogenated
solvents. Halogens are elements found in the 17th group of the periodic table.
Dueterated solvents: These solvents have deuterium a hydrogen isotope in
their molecular structure. They are preferred in experiments where hydrogen
has to be avoided. For example, in nuclear magnetic spectroscopy, the
solvents with hydrogen can interfere in the analysis. Hence, solvents
substituted with deuterium instead of the hydrogen atom are preferred. Their
examples include
Deuterated water (D2O), Deuterated methanol (CD3OD), Deuterated acetic
acid (CD3COOD), Deuterated triflouro acetic acid (CF3COOD) etc.

Based on their behavior and properties, solvents are selected for purposes
like acid-base titration, complexometry, extraction procedures, solubilization,
chromatography, spectrophotometry, etc.

The above nature seems highly specific. Because sugar (C12H22O12)

molecules appear to be organic by nature due to the presence of carbon in it.
But interestingly sugar is insoluble in organic solvents like benzene. This is
because sugar molecules have polarity and require polar solvents to dissolve.
Hence we see sugar dissolves well in plain water which is inorganic but
having polarity.

So among the types of solvents available, to dissolve a solute, one should

consider both chemistry and also the polarity.

NMR Chemical shifts

 The chemical shift is the position on the δ scale (in ppm) where the
peak occurs.
 Typical δ /ppm values for protons in different chemical environments
are shown in the schematic figure below.
 There are two major factors that influence chemical shifts

(a) deshielding due to reduced electron density (due electronegative

atoms) and
(b) anisotropy (due to magnetic fields generated by π bonds).

Note that the figure shows the typical chemical shifts for protons being
influenced by a single group.
In cases where a proton is influenced by more than one group, the effects
are essentially cumulative.

Coupling in H-NMR
So far the H-NMR spectra that we have looked at have all had different
types of protons that are seen as singlets in the spectra. This is not the
normal case.... spectra usually have peaks that appear as groups of peaks
due to coupling with neighbouring protons, for example, see the spectra of
1,1-dichloroethane shown below.

Before we look at the coupling, lets review the assignment of the peaks
first:

 δ = 5.9 ppm, integration = 1H deshielded : agrees with the -

CHCl2 unit
 δ = 2.1 ppm, integration = 3H : agrees with -CH3 unit

Now, what about the coupling patterns ?

Coupling arises because the magnetic field of vicinal (adjacent) protons
influences the field that the proton experiences.

To understand the implications of this we should first consider the effect the -CH group has
on the adjacent -CH3.

The methine -CH can

adopt two alignments
with respect to the
applied field. As a
result, the signal for
the adjacent methyl -
CH3 is split in two
lines, of equal
intensity, a doublet.

Now consider the effect of the -CH3 group has on the adjacent -CH .
The methyl -
CH3 protons give rise
to 8 possible
combinations with
respect to the applied
field. However, some
combinations are
equivalent and there
are four magnetically
different effects. As a
result, the signal for
the adjacent methine
-CH is split into four
lines, of intensity
ratio 1:3:3:1,
a quartet.

 The proximity of "n" equivalent H on neighbouring carbon atoms,

causes the signals to be split into "n+1" lines.
 This is known as the multiplicity or splitting or coupling pattern of
each signal.
 Equivalent protons (or those with the same chemical shift) do not
show coupling to each other.
 If the neighbours are not all equivalent, more complex patterns arise
(this is because of different J values, see below).
 When looking at H-NMR, the most common couplings that are
observed are those between H atoms on neighbouring C atoms (i.e.
vicinal coupling, Ha-C-C-Hb where 3 bonds are involved)
 To a first approximation, protons on adjacent sp3 C tend to behave as
if they are equivalent (for example, the H-NMR of 1-bromopropane)
Now we can do more a complete analysis, including the application of the
"n+1" rule to 1,1-dichloroethane:

 δ = 5.9 ppm, quartet, integration = 1H, deshielded : agrees with the -

CHCl2 unit next to a -CH3 unit (n = 3, so n + 1 = 4 lines)
 δ = 2.1 ppm, doublet, integration = 3H : agrees with -CH3 unit, next to
a -CH- (n = 1, so n + 1 = 2 lines)

Coupling Constant, J

The coupling constant, J (usually in frequency units, Hz) is a measure

of the interaction between a pair of protons.
In a vicinal system of the general type, Ha-C-C-Hb then the coupling
of Ha with Hb, Jab, MUST BE EQUAL to the coupling
of Hb with Ha, Jba, therefore Jab = Jba.
The implications are that the spacing between the lines in the coupling
patterns are the same as can be seen in the coupling patterns from the
H-NMR spectra of 1,1-dichloroethane (see left).

Typical Coupling Constants

So far we have emphasised vicinal coupling of H atoms on adjacent sp3 C
atoms. This coupling constant is typically about 6-8 Hz. Coupling is
controlled by geometry and the orbitals involved between the coupling
nuclei and therefore other types of systems have slightly different coupling
constants as shown below:
Notes:

 H atoms on a CH2 of a alkene don't have to be equivalent (depending

on what substituents are on the other end of the alkene). As a result
they can couple to each other. This is a 2 bond coupling.
 Coupling between H atoms that are more than 3 bonds is also
possible and is known as "long range coupling".
 In general, the more bonds involved between the H that are coupling,
the smaller the J value.
 Long range coupling is more common with rigid systems including
the pi bond systems of alkenes and substituted benzenes.
 The presence of different couplings (i.e. different J values) tends to
lead to "complex coupling" (more on that later).

Pascal's Triangle
The relative intensitites of the lines in a
coupling pattern is given by a binomial
expansion or more conviently by Pascal's
triangle.
To derive Pascal's triangle, start at the apex,
and generate each lower row by creating
each number by adding the two numbers
above and to either side in the row above
together. The first six rows are shown to the
right.
So for H-NMR a proton with zero
neighbours, n = 0, appears as a single line, a
proton with one neighbours, n =1 as two
lines of equal intensity, a proton with two
neighbours, n = 2, as three lines of intensities
1 : 2 : 1, etc.
C-NMR Spectroscopy
It is useful to compare and contrast H-NMR and C-NMR as there are
certain differences and similarities:
13
 C has only about 1.1% natural abundance (of carbon atoms)
12
 C does not exhibit NMR behaviour (I=0)
13
 C nucleus is also a spin 1/2 nucleus
13
 C nucleus is about 400 times less sensitive than H nucleus to the
NMR phenomena
 Due to the low abundance, we do not usually see 13C-13C coupling
 Chemical shift range is normally 0 to 220 ppm
 Chemical shifts are also measured with respect to tetramethylsilane,
(CH3)4Si (i.e. TMS)
 Similar factors affect the chemical shifts in 13C as seen for H-NMR
 Long relaxation times (excited state to ground state) mean no
integrations
 "Normal" 13C spectra are "broadband, proton decoupled" so the
peaks show as single lines
 Number of peaks indicates the number of types of C

The general implications of these points are that 13C-NMR spectra take
longer to acquire than H-NMR, though they tend to look simpler. Accidental
overlap of peaks is much less common than for H-NMR which makes it
easier to determine how many types of C are present.

What does broadband, proton decoupled mean ?

The resonances due to 13C nuclei are split by neighbouring H atoms.

These splittings would complicate the appearance of the spectra making
them harder to interpret. Therefore, in a "normal" 13C spectra, these
couplings are "removed" by applying a continuous second radio frequency
signal of a broad frequency range that excites all the H nuclei and cancels
out the coupling patterns due to the interaction of the H with the 13C. This
means that each C is seen as a single line. Of course information is being
lost by doing this, such as how many H are attached to each C.

In off-resonance decoupling the one bond C-H couplings are retained so

the signal for a particular C is given by the number of attached H in accord
with n+1 rule. So, for example, a -CH3 shows as a quartet and a -CH2- as a
triplet.

QUESTIONS
  How many lines in the peak for a C-H (i.e. methine) group in an off-
resonance decoupled spectra ? ANSWER
 What about a quaternary C with no H attached ? ANSWER

13
C chemical shifts
The most significant factors affecting the chemical shifts are:

 Electronegativity of the groups attached to the C

 Hybridisation of C

A simple correlation table of 13C chemical shifts is shown below.

2D-NMR Spectra
 So far the NMR spectral methods we have discussed have been one
dimensional (since they have a single chemical shift x coordinate
axis).
 With the development of more advanced spectroscopic methods as
computational power has increased, it has become possible to obtain
two dimensional spectra.
 In two dimensional experiments, both the x and the y axes have
chemical shift scales and the 2D spectra are plotted as a grid like a
map.
 Information is obtained from the spectra by looking at the peaks in
the grid and matching them to the x and y axes.

 COSY - Correlation Spectroscopy

o both axes correspond to the proton nmr spectra.
 o the COSY spectra indicates which H atoms are coupling with
each other.
o An example of a COSY is provided below.

 HETCOR - Heteronuclear Correlation Spectroscopy

o proton nmr spectra on one axis and the 13C nmr spectra on
the other.
o the HETCOR spectra matches the H to the appropriate C.

COSY spectra

 The information on the H that are coupling with each other is

obtained by looking at the peaks inside the grid. These peaks are
usually shown in a contour type format, like height intervals on a
map.
 In order to see where this information comes from, let's consider an
example shown below, the COSY of ethyl 2-

butenoate
 First look at the peak marked A in the top left corner. This peak
indicates a coupling interaction between the H at 6.9 ppm and the H
at 1.8 ppm. This corresponds to the coupling of the CH3 group and
the adjacent H on the alkene.
 Similarly, the peak marked B indicates a coupling interaction
between the H at 4.15 ppm and the H at 1.25 ppm. This corresponds
to the coupling of the CH2 and the CH3 in the ethyl group.
 Notice that there are a second set of equivalent peaks, also
marked A and Bon the other side of the diagonal.
 (COSY spectra recorded by D. Fox, Dept of Chemistry, University of Calgary on a Bruker Advance DRX-400 spectrometer)

HETCOR spectra

 The information on how the H are C are matched is obtained by

looking at the peaks inside the grid. Again, these peaks are usually
shown in a contour type format, like height intervals on a map.
 In order to see where this information comes from, let's consider an
example shown below, the HETCOR of ethyl 2-butenoate.
 First look at the peak marked A near the middle of the grid. This
peak indicates that the H at 4.1 ppm is attached to the C at 60
ppm. This corresponds to the -OCH2- group.
 Similarly, the peak marked B towards the top right in the grid
indicates that the H at 1.85 ppm is attached to the C at17
ppm. Since the H is a singlet, we know that this corresponds to the
CH3- group attached to the carbonyl in the acid part of the ester
and not the CH3- group attached to the -CH2- in the alcohol part of
the ester.
 Notice that the carbonyl group from the ester has no "match" since it
has no H attached in this example.