13.

Nuclear Magnetic Resonance (NMR)

Spectroscopy
A. Basic Principles
Nuclear magnetic resonance (NMR) spectroscopy is one of the most important and widely used
methods for determining the structure of organic molecules. NMR allows one to deduce the
carbon-hydrogen connectivity in a molecule. The following experiment provides a general
overview of NMR theory and interpretation. For a more extensive discussion, you should consult
your textbook.

The nucleus of a hydrogen atom (1H) as well as certain other nuclei (13C, 15N, 31P, etc.) possess
a property called nuclear spin. Any nucleus that has an odd mass number and/or an odd atomic
number has a nuclear spin. Nuclear spins are oriented randomly in space, however, in the
presence of an external magnetic field (Bext) the spins align. Some spins align parallel to Bext (α
spin state) while others align antiparallel to Bext (β spin state). The nuclear spins that align
parallel to Bext are slightly lower in energy. (Figure 1)

H H H β-state
H H
B ext

H
H
H H H H H α-state

B ext

Figure 1. Nuclear Spins in the Presence of Bext

When the nuclear spins in the α and β states are irradiated with electromagnetic energy from
the radio frequency (RF), energy is absorbed and some of the nuclear spins in the α state are
“spin-flipped” to the higher energy β state. At this stage, the nuclei are said to be in resonance
with the applied RF radiation. (Figure 2)

H H β-state H H H
RF
Radiation

H H H H α-state H H H
B ext B ext
Nuclei are in resonance
with the RF radiation

Figure 2. Nuclei Irradiated with RF Radiation

Experiment 13 – NMR Spectroscopy Page 1 of 10

 Over time, the flipped spins slowly fall back down to the higher energy state. The signal decay is
detected over time producing a raw signal called a free induction decay (FID). A mathematical
function called Fourier Transform converts the FID into the NMR spectrum. (Figure 3)

H H H H H
Relaxation

H H H H H H H
B ext B ext

1H NMR Spectrum

Fourier
Transform

FID 3 2 1 0
PPM

Figure 3. Relaxation and Generation of the NMR Spectrum

Every distinct proton in a molecule will require a specific RF frequency to promote the “spin-flip.”
Thus, the energy difference between the α and β states for each proton is distinct. Depending
on the energy difference (ΔE), the absorption of each proton on the NMR spectrum will appear
at different locations.

B. The NMR Instrument

The NMR spectrometer contains a very strong magnet, which
generates an extremely strong, uniform magnetic field. The magnet
poses no health hazard, however, you should not bring certain
items such as watches, credit cards, and cellphones close to it. The
NMR instrument in our laboratory is a 400 MHz instrument, which
corresponds to a magnetic field of 9.4 Tesla. A higher magnetic field
will produce a higher resolution NMR spectrum, however, the
instrument cost rises steeply with magnetic field strength.

The sample is placed in an NMR tube, which is inserted into the

center of the instrument. The tube is spun rapidly to improve
homogeneity resulting in a better spectrum. A radio frequency (RF)
console is attached to the spectrometer; this console sends an
intense pulse of RF energy to the sample. The RF pulse covers a
wide enough range to excite all of the protons in the molecule. A
computer collects the resulting FID and converts it to the NMR
spectrum, which can be printed and analyzed.
NMR Spectrometer

Experiment 13 – NMR Spectroscopy Page 2 of 10

 C. Sample Preparation
NMR samples are typically prepared as a dilute solution in a deuterated solvent. A
typical sample size is 5-10 mg of sample dissolved in 0.5 – 1.0 mL of deuterated
solvent. Less than 1 mg of sample can be used, however, this will necessitate a
longer NMR acquisition time. Deuterated solvents are ones in which all of the
hydrogen in the molecule have been replaced with the deuterium isotope. Deuterium
does not show up in the range examined for 1H NMR, thus the deuterated solvent is
effectively NMR transparent. Some common NMR solvents are shown below. The
most commonly used NMR solvent is chloroform-d, where the single hydrogen in
chloroform is replaced by deuterium.
O

CDCl 3 D 3C CD 3 D 2O

chloroform-d acetone-d6 deuterium oxide

O
S
D 3C OD D 3C CD 3

methanol-d 4 DMSO-d 6
Common NMR Solvents
Sometimes, an NMR solvent is spiked with tetramethylsilane (TMS), which
NMR Tube
provides a reference signal that can be used to calibrate the spectrum. The TMS
signal is set at exactly 0 ppm and every other signal is based off of that
reference point.

NMR samples are placed inside a specially designed NMR tube, which is a 5 mm precision
glass tube that fits perfectly inside the instrument. Once inside the tube, the diluted sample
should have a height of 2-4 cm. NMR tubes are reusable and should be washed out with
acetone and left to dry thoroughly before the next use.

Great care should be taken when inserting the sample tube into the spectrometer. If a sample
tube breaks inside the instrument, a great deal of time is required to remove the broken glass
and to get the instrument back up and running.

D. Interpretation of NMR Spectra

1. Basics of the 1H NMR Spectrum
The interpretation of 1H NMR spectra involves three basic concepts: chemical shift, peak
integration, and coupling. Each of these items provides important information regarding the
molecular structure. Combined, this information can be used to elucidate the structure of an
unknown organic molecule.

Figure 4 shows a sample 1H NMR spectrum. The x-axis is the chemical shift (δ) expressed in
parts per million (ppm). The typical chemical shift scale for proton NMR is 0-12 ppm. The
chemical shift provides information regarding the chemical environment of the corresponding
proton. The area under each peak is know as the integration. The integration for a signal is

Experiment 13 – NMR Spectroscopy Page 3 of 10

 proportional to the number of hydrogen corresponding to that particular signal. Finally, you will
notice that some peaks are split (b and c, figure 4). Peak splitting is also known as coupling.
The splitting for an individual signal provides information as to the number of hydrogen present
on the adjacent carbon atoms.
b c
H2 a
c C OH
H 3C

O
b

10 8 6 4 2 0
PPM

Figure 4. Sample Proton NMR Spectrum

2. Chemically Equivalent and Distinct Hydrogen

Every chemically distinct type of hydrogen in a molecule gives rise to an individual signal in the
proton NMR spectrum. This is in contrast to chemically equivalent hydrogen, which cannot be
differentiated and will be represented by a single signal. For example, methane (figure 5A) has
four protons. These four protons cannot be distinguished and are thus all chemically equivalent.
Since there is only one type of hydrogen in methane, only one proton NMR signal will result. In
ethanol (figure 5B), there are three distinct types of hydrogen: the OH proton, the CH2 group,
and the CH3 group. Note that the two protons in the CH2 group are equivalent and the three
protons in the CH3 group are equivalent. When analyzing a molecule for chemically equivalent
or distinct atoms, pay special attention to molecular symmetry. Butane (figure 5D), for example,
has a plane of symmetry, thus there are two equivalent ethyl (CH2CH3) groups in the molecule.
b b b
H H2 a H2 a H2
a C H 3C C H 3C C a
H C H c c
HO CH 3 O CH 3 C CH 3
H b H2
1 type of H 3 types of H 3 types of H 2 types of H
1 NMR Signal 3 NMR Signals 3 NMR Signals 2 NMR Signals
(A) (B) (C) (D)

Figure 5. Examples of Chemically Equivalent and Distinct Protons

Chiral Centers and Diastereotopic Protons
In an achiral molecule, the two hydrogen in a methylene -CH2- group are typically chemically
equivalent (see figure 5 B-D). If however, a chiral center (*) is present, the two methylene
protons are no longer equivalent and will each have its own distinct NMR signal. In this type of
situation, the methylene protons are said to be diastereotopic. For example, the molecule in
figure 6A does not have a chiral center and the two highlighted hydrogen of the CH2 group are
chemically equivalent. The molecule in figure 6B has a chiral center, which makes the two
hydrogen in the CH2 group diastereotopic and chemically distinct. One special case is that
shown in figure 6C where there is not a chiral center, yet the two hydrogen in each methylene
group are diastereotopic and chemically distinct. This can be rationalized by thinking three
dimensionally. Considering group Z as an “out” group on a wedge, reveals the spatial difference
between each methylene hydrogen and Z. One hydrogen is “out” in a closer spatial orientation

Experiment 13 – NMR Spectroscopy Page 4 of 10

 to Z, while the other hydrogen is “back” and is further away from Z. These two hydrogen reside
in different chemical environments and will have different NMR shifts. Diastereotopic hydrogen
give rise to complex NMR signals that can often be difficult to interpret.

Z Z
H H H H
O * O H HH H H H

Chemically Equivalent Chemically Distinct Chemically Distinct

CH2 protons CH2 protons CH2 protons
(A) (B) (C)

Figure 6. Examples of Diastereotopic Protons

3. Chemical Shift
The chemical shift of a proton gives us insight into the chemical environment of that proton – i.e.
what type of groups are nearby. Chemical shift increases from right to left on the NMR
spectrum. The right end of the spectrum is considered upfieled and protons in this region are
said to be shielded. The left end of the spectrum is considered downfield and protons in this
region are said to be deshielded. (Figure 7)
downfield upfield
deshielded protons shielded protons

12 10 8 6 4 2 0
PPM
ppm

Figure 7. Chemical Shift Terminology

All proton NMR chemical shifts are relative to the signal produced by tetramethylsilane (TMS),
which has a chemical shift that is defined at δ = 0 ppm. TMS is often used as an internal
standard and the spectrum can be referenced to the chemical shift of this standard.

Chemical shifts are remarkably predictable and knowing a few base values along with some
simple trends will allow you analyze a variety of NMR spectra. Some typical proton NMR regions
are shown in figure 8.
Alcohol -OH / Amine -NH 2
Z R-CH 3
H ~0.9
O O H
H H ~1.2
Z= O,C
OH H
H H
carboxylic aldehyde aromatic vinyl H ~1.7
acid X
Csp3-H
X = O, N, halogen saturated

12 10 8 6 4 2 0
PPM

Figure 8. Typical 1H NMR Chemical Shift Regions

Experiment 13 – NMR Spectroscopy Page 5 of 10

 Chemical Shift Origin
Examination of figure 8 reveals that protons in different chemical environments display NMR
resonances (signals) at different chemical shifts. The protons in a molecule are each shielded
by the nearby electrons. In the presence of an external magnetic field (Bext) the molecule’s
electrons produce small local magnetic fields (Blocal) that opposes the applied field from the
instrument. In effect, the proton is shielded from the external magnetic field by the opposing
local magnetic field. The overall magnetic field experienced by an individual proton is the
effective magnetic field (Beffective) A smaller Beffective, means that due to shielding, a proton
experiences less of the external magnetic field resulting in a lower chemical shift.
Beffective = Bext - Blocal
Protons in more electron deficient environments are less shielded from the external magnetic
field and resonate at higher chemical shifts. Protons at higher chemical shifts are said to be
deshielded.

Chemical Shift Predictions

Chemical shift values are typically additive. Starting with a base value, a chemical shift value
can be approximated by incrementally adding the chemical shift effect from each attached
group. Base values for the methyl, methylene, and methine groups are shown below.

Methyl Methylene Methine

H H
C H C C H
H H
~0.9 ppm ~1.2 ppm ~1.7 ppm

For example, adding an electronegative chlorine increases a proton’s chemical shift by ~2.2
ppm. Thus, chloromethane has proton NMR signal at 0.9 + 2.2 = 3.1 ppm. Chloroethane, on the
other hand, has a proton NMR signal at 1.2 + 2.2 = 3.4 ppm corresponding to the methylene
group.
Cl CH 3 Cl C CH 3
H2

3.1 ppm
3.4 ppm
The presence of one or more electronegative atoms will deshield a nearby proton by the
inductive effect. The more electronegative the attached atom, the stronger the deshielding effect
and the higher the chemical shift. Some representative examples are shown below.

H H H H H
O C H Cl C H Br C H I C H H 2N C H
H H H H H
δ ~3.4 ppm 3.1 ppm 2.7 ppm 2.2 ppm 2.4 ppm
e- neg O – 3.5 Cl – 3.0 Br – 2.8 I – 2.7 N – 3.0

Additionally, as the number of electronegative atoms near a particular proton increases, the
chemical shift also increases.

CH3CH3 CH3Cl CH2Cl2 CHCl3

0.9 ppm 3.1 ppm 5.3 ppm 7.3 ppm

Experiment 13 – NMR Spectroscopy Page 6 of 10

 Alcohol and Amine Protons
The OH group of an alcohol and NH/NH2 group of an amine typically gives a broad singlet in the
0-5 ppm range. In most cases, OH and NH protons are not split by protons on neighboring
atoms.

Alkene and Aromatic Protons

Protons with nearby π-electrons, specifically vinyl and aryl protons, have unusually high
chemical shifts. This is due to an effect known as magnetic anisotropy. In these systems, the
local magnetic field aligns with the external magnetic field, causing the proton to have a larger
than expected effective magnetic field.

Vinylic protons typically appear in the 4.5-6.5 ppm region. Terminal alkene protons tend to have
a slightly lower chemical shift than an internal alkene proton (4.9 vs 5.4 ppm).

Aryl protons typically appear in the 6.5-8 ppm region. A phenyl group –C6H5 has three distinct
types of protons, however, the signals for these protons frequently overlap and show up as a
single line corresponding to 5 hydrogen or a messy grouping of peaks corresponding to 5
hydrogen. When the aromatic ring is di or tri substituted, the aryl proton signals are typically
better resolved, especially when one of the substituents is an electronegative atom or electron
withdrawing group.

4. Integration
It is important to recognize that the area under an NMR signal is proportional to the number of
protons corresponding to that particular signal. Modern NMR spectrometers have the ability to
automatically integrate each signal providing relative peak areas with ease.

For example, the integrated NMR spectrum of 2-butanone is shown in figure 9. The area under
each integral is proportional to the number of H represented by that particular signal. Methyl
group a, has three equivalent hydrogen. This is represented in the NMR spectrum by an
integration for 3H. The methylene group b has two equivalent hydrogen and therefore integrates
for 2H. Finally, methyl group c has three equivalent hydrogen and integrates for 3H.

c
a b

a c
b 3H
3H
2H

3 2 1 0
PPM

Figure 9. NMR Spectrum of 2-Butanone

In some cases, you will have integration values that need to be normalized based on the
smallest integration value. This process is further explored in pre-lab question 4.

Experiment 13 – NMR Spectroscopy Page 7 of 10

 5. Spin-Spin Splitting (Coupling)
The signal corresponding to a particular proton will split as a result of the protons on neighboring
carbon atoms. Basic splitting follows the n+1 rule where n is the number of protons on the
adjacent carbon.

Looking at the NMR spectrum of 2-butanone (figure 9), you will see that signal b is split into four
lines and signal c is split into three lines. The reason for this observed splitting is due to
coupling with the protons on the adjacent carbon atoms. Figure 10 shows 2-butanone along with
the observed splitting for each proton in the molecule. The signal corresponding to a appears as
a single line (singlet). This is because it is not coupled to any other hydrogen. The adjacent
carbon does not have any hydrogen for a to couple with. The signal corresponding to b on the
other hand is split into four lines (quartet). This is due to one of the adjacent carbons having
three protons attached. Following the n + 1 rule, with n = 3, 3 + 1 = 4 (quartet). Finally, the
signal corresponding to c is a triplet because the adjacent carbon has two protons (2 + 1 = 3).

O O O
c
a b

No H on the adjacent C 3 H on the adjacent C

2 H on the adjacent C
No splitting n +1 = 3 +1 = 4
Single Line = singlet Split into four lines = quartet n +1 = 2+1 = 3
Split into three lines = triplet

Figure 10. Splitting in 2-Butanone

The most common splitting patterns, also known as multiplicities, are shown in figure 11. Once
a peak is split into more than seven lines, it is commonly referred to as a multiplet, m.

Singlet Doublet Triplet Quartet Quintet Sextet Septet

s d t q quin. sex. sep.
Figure 11. Common Multiplicities

Complex Splitting
When a proton is coupled to two or more sets of not equivalent hydrogen, the splitting becomes
more complex and follows the (m + 1)(n + 1) splitting pattern. Consider for example the alkene
shown in figure 12a. The carbon highlighted in the green box is coupled with two different types
of hydrogen. Coupling with the alkene H, splits the signal into a doublet. Coupling with the CH2
hydrogens further splits the signal into a triplet providing a doublet of triplets. A splitting tree
Experiment 13 – NMR Spectroscopy Page 8 of 10
 (figure 12b) is used to show how the signal splits into a double of triplets. Think of the initial
signal first splitting into two (doublet) then each of these two lines further splitting into three lines
(triplet). In reality, the peaks often overlap which results in the observation of fewer than
expected peaks (figure 12c).

doublet
H 3C CH 3

H2 m = 2
C triplets
n =1 H C CH 3
H2
2+1 = 3
1 +1 =2
(2)(3) = doublet of triplets

Ideal: Well Reality: peaks

separated d of t often overlap

(a) (b) (c)

Figure 12: Complex Splitting

Coupling Summary
Several important features regarding coupling are listed below.
1. The most common coupling occurs between non-equivalent protons that are separated by
three bonds.
H O
Br
Cl Cl
Br H
Cl
H H
2. Coupling will be observed between protons separated by two bonds if the two protons are
chemically distinct.
H
Each alkene hydrogen appears as a doublet because
they are chemically distinct and coupled to one another.
H Cl
3. Coupling is typically not observed through heteroatoms such as oxygen and nitrogen.
H H Singlet
Singlet H
O Cl
4. Chemically equivalent protons do not couple.
H H
Cl Both CH2 groups are chemically equivalent.
Cl They show up as a singlet integrating for 4H
H H

The Coupling Constant (J – Values)

The coupling constant (aka J - value) is the distance between two peaks in a split signal. If two
protons are coupled to one another, they will have identical coupling constants. When analyzing
complex spectra, the coupling constant is a convenient piece of data to help determine whether
or not two signals are coupled to one another. The coupling constant for a particular signal is
calculated by subtracting the chemical shift of two neighboring peaks in a signal and multiplying
that number by the frequency of the spectrometer. The NMR below was taken on a 300 MHz
instrument. The coupling constant of the quartet at ~3.4 ppm is: (3.48 – 3.45) x 300 = 9 Hz. Now

Experiment 13 – NMR Spectroscopy Page 9 of 10

 prove that this quartet is coupled to the triplet by calculating its coupling constant. Tables of
common coupling constants can be found in most organic textbooks and online.

1.49 ppm
H2
C
Cl CH 3

3.42 ppm 1.52 ppm

3.45 ppm 1.46 ppm

3.48 ppm 3.39 ppm

4 3 2 1 0
PPM

E. Experimental Procedure
A set of unknown IR and NMR spectra for a variety of organic compounds will be available in
the laboratory. A list of possible structures will be provided separately. Your TA will assign you
two of the numbered unknowns. You will analyze the NMR data and match your spectra to one
of the possible structures. On the report sheet, you should identify each of your unknowns. You
should include a list of the pertinent IR absorptions and NMR signals along with the identity of
the group that each signal corresponds to. You should also include a brief explanation of how
you identified your unknown based on piecing together this data.

Once you have completed your unknown identification you are encouraged to use the remaining
lab time analyzing other NMR spectra for additional practice.

Experiment 13 – NMR Spectroscopy Page 10 of 10