Study materials for the final exam

An introduction to spectroscopic analysis

methods.

March 23rd, 2024

Contents

An introduction to spectroscopic analysis methods 1

Basics 1
IR Spectroscopy 1
1H-NMR Spectroscopy 3
13C-NMR Spectroscopy 9

NMR Spectroscopy Tasks 11

NMR Spectroscopy Tasks – Solutions 15

An introduction to spectroscopic analysis methods
I. Basics
𝐸 =ℎ·𝜈
Molecules can assume different energy levels (e.g. vibrational states, electrons in differ-
ent orbitals, …). Transitions between the different states are quantised, i.e. bonds can e.g.
only be stretched or twisted with certain frequencies and electrons can only jump be-
tween orbitals with defined energy differences. We measure these energy differences
𝐸2 −𝐸1
𝐸2 − 𝐸1 or frequency differences 𝜈 = in the form of different types of spectra.
ℎ

II. IR Spectroscopy
Absorption of infrared light changes the vibrations in a molecule. Covalent bonds are
stretched (stretching vibrations) or rotated around bond angles = "deformed" (defor-
mation vibrations). The corresponding absorption bands occur in sharply defined wave-
length ranges, depending on the bond strength and mass of the bound atoms. IR spectros-
copy is therefore particularly suitable for the detection of bond types (functional groups)
in a molecule. IR frequencies are usually given as wavenumbers, defined as the number
of vibrations per cm (measuring range 4000 to 600 cm-1).
𝜈
Wavenumber 𝜈̃ = 𝑐

The characteristic bands of functional groups always appear in the same range, regard-
less of the exact molecular structure. Typical wavenumbers for stretching vibrations of
functional groups:

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Example: Differentiation between the isomeric structures C6H5CH2OH and C6H5OCH3

Look for characteristic IR bands of the compound classes from tables. In this case, the
benzyl alcohol should have a strong, broad peak in the range 3400-3700 cm-1 for the OH
stretching vibration, but the isomeric ether should not.

Deformation vibrations are excited by low frequency light. The corresponding absorption
bands appear as a complicated pattern in the range between 1500 and 600 cm-1. This so-
called fingerprint region is normally not interpreted individually, but is characteristic for
certain compounds.

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 Crash course
on youtube!
III. 1H-NMR-Spectroscopy

Nuclear magnetic resonance spectroscopy is a modern analytical method for

the structural elucidation of molecules.

NMR spectroscopy makes use of the fact that not only electrons, but also certain atomic
nuclei have a non-zero spin, the nuclear spin. These include the nuclei of the nuclides 1H
and 13C. If a sample of a compound is brought into a strong, homogeneous magnetic field,
these nuclei behave like tiny magnets and align themselves in the magnetic field. The nu-
clear spins thereby occupy a low energy level. If the system of aligned nuclear spins is
disturbed by the irradiation of electromagnetic energy in the radio wave range (wave
length 0.5 - 5 m), then the nuclear spins can be raised to a higher energy level, for example
by flipping in the magnetic field. For a given nucleus, the energy required for this essen-
tially depends on the strength of the external magnetic field. Today's measuring instru-
ments, which use magnetic fields in the range of 1.4 to 14 Tesla, emit frequencies of 60 to
900 MHz. To a certain extent, the required energy also depends on the environment of
the nuclei in the molecule. With the help of this dependence, information can be obtained
about the structure of the compound under investigation!

To record a 1H NMR spectrum, the sample is dissolved in a solvent that has no 1H nuclei,
such as CCl4 or CDCl3. The reference substance tetramethylsilane (TMS, Si(CH3)4) is also
added. The glass tube with the sample is placed in the middle of the high-frequency coil
between the poles of a strong magnet. The nuclei immediately orient themselves parallel
or antiparallel to the field. A continuously increasing energy can be applied to the nuclei
via the high-frequency coil. If the radiated energy corresponds exactly to the energy dif-
ference between the lower-energy and the higher-energy spin state, it is absorbed by the
nuclei (i.e. the nuclei are in resonance with the radiation). The energy absorbed by the
sample is plotted as a function of the applied frequency.

For the interpretation of an NMR spectrum, 3 main points are taken into account:

1. Chemical Shift

It is common to plot the frequency of the radiated energy in ppm (parts per million, i.e. in
units of 10-6) of the absolute amount. This means that for a single-beam frequency of 200
MHz, 1 ppm corresponds to 200 Hz. This has the advantage that the numerical values
become independent of the magnetic field strength of the spectrometer used.

The resonance frequencies of the 1H nuclei of the investigated compounds appear as a so-
called chemical shift in ppm, relative to the added reference substance TMS (=zero
point).

The chemical shift of hydrogen nuclei in organic molecules is influenced by the type of
functional groups. Depending on the structure of the molecule, the chemical shifts of iden-
tical functional groups differ slightly from each other, so that the NMR spectrum is char-
acteristic for a substance.

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Two factors in particular are important for the chemical shift:

a) The electronegativity of the atoms or groups of atoms in the immediate vicinity to

the proton. Electron-withdrawing groups (e.g. halogens, oxygen, ...) generally
cause a low-field shift (further away from the TMS standard).
b) -Electrons
e.g.: -CH3 (at 0,9), -CH2Cl (at 3,7), -CHCl2 (at 5.8)
e.g.: -C-CH2- (at 1,2-1,4), CH2=CH- (at 4,6-5,0), H-C6H5 (t 6,6-8,0)

Example: 1,4-Dimethylbenzol

 = 0 ppm: TMS Reference

 = 2,30 ppm: 6 1H-nuclei of the methyl groups

 = 7,10 ppm: 4 1H-nuclei of the aromatic

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 2. Signal surfaces

The area under each signal is determined by integration. The signal area is directly pro-
portional to the number of 1H nuclei (integration lines are determined by the meter and
printed out directly). In the example above, the ratio is 42:29 = approx. 3:2 (or 6:4 if the
sum formula is known). This allows the assignment of the signal at = 2.30 ppm to the six
methyl protons and the signal at = 7.10 ppm to the equivalent aromatic protons.

3. Spin-spin coupling

Many compounds provide spectra that contain not only single signals (singletons), but
complicated signal patterns (multipletons) for the different types of protons. In the so-
called spin-spin coupling, the 1H nucleus, in addition to the external magnetic field, also
"feels" the weak fields of the neighbouring 1H nuclei. These tiny fields "disturb" the mag-
netic field of the nucleus whose signal we are observing.

The splitting is done according to the (n+1) rule: If a 1H nucleus has n 1H neighbours of
different chemical shift, its signal splits into (n+1) signals. Accordingly, the spin-spin cou-
pling pattern of the signals yields the number of nearest 1H neighbours of a certain type
of 1H nuclei.

Examples:

Ethanol:

𝛿 = 1,10: triplet signal

The three identical 1H nuclei belonging to the CH3 group ‘see’ two neighbouring 1H nuclei,
which gives a (2+1)= 3 (triplet) signal.

𝛿 = 3,60: quadruplet signal

The two identical 1H nuclei belonging to the CH2 group ‘see’ three neighbouring 1H nuclei
(belonging to the CH3 group), which gives a (3+1) = 4 (quadruplet) signal.

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This signal is shifted further down field (𝛿 = 3,60) due to the proximity of the O-atom
(c.f. table page 9).

Note: spin-spin coupling usually only occurs between 1H-nuclei of neighbouring C-atoms,
which is why the 1H-nucleus of the alcohol group (𝛿 = 3,70, broad signal) shows no cou-
pling pattern in this example.

Dimethylether:

The 1H nuclei can’t ‘see’ any direct neighbours, so no spin-spin coupling occurs. As the
molecule is perfectly symmetric, the 1H nuclei from both methyl groups give the same
singlet signal.

Diethylether

𝛿 = 1,10: triplet signal

The three identical 1H nuclei belonging to the CH3 group ‘see’ two neighbouring 1H nuclei,
which gives a (2+1)= 3 (triplet) signal.

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𝛿 = 3,50: quadruplet signal

The two identical 1H nuclei belonging to the CH2 group ‘see’ three neighbouring 1H nuclei
(belonging to the CH3 group), which gives a (3+1)= 4 (quadruplet) signal.

This signal is shifted further down field (𝛿 = 3,50) due to the proximity of the O-atom
(c.f. table page 9).

As the molecule is perfectly symmetric, the 1H nuclei from both CH3 and CH2 groups give
the same signals.

Benzene

𝛿 = 7,30: singlet signal

All six 1H nuclei are chemically identical, as the molecule is perfectly symmetric. They all
give the exact same signal, which gives 1 singlet signal at 𝛿 = 7,30.

Methyl benzoate

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𝛿 = 3,90: singlet signal

The three identical 1H nuclei belonging to the CH3 group don’t ‘see’ any neighbouring 1H
nuclei, thus they give a singlet signal that is shifted down field due to the proximity to the
ester function.

𝛿 = 7,50 to 8,10: multiple signals that add up to a relative intensity of 5, which corre-
spond to the five 1H nuclei of the benzene. The signals of aromatic rings can sometimes
be split perfectly, or they may partially overlap, depending on the other functions at-
tached to the benzene ring.

Note

The distance between any two adjacent lines in the NMR peaks of two sets of equivalent
hydrogen nuclei coupled only to each other is the same, which, when expressed in hertz
(Hz) is called the coupling constant (symbol: 𝐽) of the two sets of equivalent hydrogen
nuclei.

e.g.:

To convert x, which is in ppm, into the coupling constant, multiply x by the operating fre-
quency, expressed in MHz, of the NMR spectrometer used for the experiment. The x for
two sets of equivalent hydrogen nuclei coupled to each other depends on the operating
frequency of the spectrometer, but J does not.

Coupling constants are influenced by the type of bond and take up a characteristic value,
as indicated in the following table.

H-H coupling constants (in Hz)

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 IV. 13C-NMR-Spectroscopy

The differences to 1H NMR spectroscopy are, that the chemical shifts extend over a
larger range (0 - 200 ppm, also measured against the standard TMS) and, because of
the low abundance of 13C, there are usually no spin-spin couplings to 13C nuclei. How-
ever, couplings to the 1H nuclei do occur, which is why most 13C NMR spectra are ‘pro-
ton decoupled’, thus showing only singlet signals for the various C atoms.

Due to the lack of spin-spin coupling and without any type of signal integration, it can
sometimes be tricky to assign certain signals to specific C atoms. Thus, the main use of
13C NMR spectra is to determine the number of identical C-atoms within a molecule.

Examples:

Butan-2-ol

4 sharp singlets appear. The peak of the C2 atom (with the OH group) appears at the
lowest field (δ=70 ppm).

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Benzaldehyde

B
A
C

D B
C

A sharp singlet signal at 𝛿 = 191 ppm can be attributed to the 1H nucleus of the aldehyde
function.

Due to the symmetry of the molecule, the benzene ring yields another 4 distinct signals
(A, B, C and D) instead of 6. The two signals at 𝛿 = 129 ppm and 𝛿 = 130 ppm appear to
be much higher in intensity than the other signals. This hints to the fact, that they can be
attributed to two C atoms each (2x B and 2x C), even though proper integration of the
surface areas cannot be measures in these spectra.

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NMR Spectroscopy Tasks

Task 1
Assign the signals of the 1H NMR spectrum to the indicated substance.

Note:
In these types of spectra, the signal surfaces (integration lines) are visualized by the red
lines printed above the signals. These can be measured to give information about the ratio
of 1H nuclei within the various groups:
a:b:c:d=1:1:4:6

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Task 2
Assign the correct connection to the spectra:

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Task 3
Assign the 3 spectra below to substances with the molecular formula C3H8O.

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Task 4
Assign the following isomers to the corresponding spectra.

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NMR Spectroscopy Tasks - Solutions
Task 1
𝛿 = 1,1: triplet, relative intensity = 6
→ corresponds to the 6 identical 1H atoms of the CH3 groups.
𝛿 = 2,4: quadruplet, relative intensity = 4
→ corresponds to the 4 identical 1H atoms of the CH2 groups.
𝛿 = 5,0: doublet, relative intensity = 1
→ corresponds to the 1H atom adjacent to the ethoxy groups.
𝛿 = 6,6: doublet, relative intensity = 1
→ corresponds to the 1H atom of the CHCl2 group. The proximity to the chlorine atoms explains
the deep chemical shift of 𝛿 = 6,6.

Task 2
A:
The three identical 1H nuclei of CH3 group ‘see’ one neighbouring 1H atom. This would generate a
doublet signal with a relative intensity of 3.
The 1H nucleus of the CHBr2 group ‘sees’ three neighbouring 1H atoms. This would generate a
quadruplet signal with relative intensity of 1, with a deep chemical shift due to the proximity to
two bromine atoms.
→ spectrum 2.
B:
The three identical 1H nuclei of CH3 group ‘see’ two neighbouring 1H atoms. This would generate
a triplet signal with a relative intensity of 3.
The two identical 1H nuclei of CH2 Br group ‘see’ three neighbouring 1H atoms. This would
generate a quadruplet signal with a relative intensity of 2, which would be shifted down a bit, due
to the proximity to one bromine atom.
→ spectrum 1.
C:
The two identical 1H nuclei of CH2 Br group ‘see’ one neighbouring 1H atom. This would generate
a doublet signal with a relative intensity of 2, which would be shifted down a bit, due to the
proximity to one bromine atom.
The 1H nucleus of the CHBr2 group ‘sees’ two neighbouring 1H atoms. This would generate a
triplet signal with relative intensity of 1, with a deep chemical shift due to the proximity to two
bromine atoms.
→ spectrum 3.

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Task 3

CH3 group: triplet with relative intensity 3

CH2 group (adjacent to CH3 ): sextet (3+2 neighbours) with relative intensity 2
CH2 group (adjacent to OH): triplet (no coupling with OH group) with relative intensity 2
OH group: 1 broad singlet with relative intensity 1
→ spectrum 3.

CH3 group (adjacent to CH2 ): triplet with relative intensity 3

CH2 group: quadruplet with relative intensity 2
CH3 group (adjacent to ethoxy group): singlet with relative intensity 3
→ spectrum 2.

CH3 group: doublet with relative intensity 6 (both groups are identical)
CH group: septet (due to 6 neighbouring 1H nuclei) with relative intensity 1
OH group: 1 broad singlet with relative intensity 1
→ spectrum 1.

Task 4

CH3 group: triplet with relative intensity 3

CH2 group: quadruplet with relative intensity 2
OH group: 1 broad singlet with relative intensity 1
→ spectrum 2.

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CH3 group (left): singlet with relative intensity 3
CH3 group (right): singlet with relative intensity 3.
→ spectrum 3.

Single H (left): singlet with relative intensity 1, very deep shift.

CH2 group: quadruplet with relative intensity 2
CH3 group: triplet with relative intensity 3
→ spectrum 1.

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