TECH

m o d u l a r
publisher: H. A. Neidig
· l a b o r a t o r y · p r o g r a m · i n · c h e m i s t r y
organic editor: Joe Jeffers
710

Identifying an Unknown
Compound by Infrared
Spectroscopy
prepared by Moses Lee, Furman University

PURPOSE OF THE Prepare IR samples. Obtain and interpret IR spectra of organic

EXPERIMENT compounds.

BACKGROUND REQUIRED You should have experience in IR spectroscopy, including theory and
interpretation.

BACKGROUND Infrared (IR) radiation is radiation in the energy range between the
INFORMATION visible and microwave regions of the electromagnetic spectrum. The
portion of the IR spectral region between 4000 and 400 cm –1 is of
greatest practical use to the organic chemist. An IR spectrum, as
shown in Figure 1, is a plot of the percentage of IR radiation that
passes through a sample versus the frequency of the radiation. The
radiation that passes through the sample is measured as percent
transmission. The frequency of the radiation is measured in wave-
lengths per centimeter, which is known as a wavenumber.

Figure 1 An infrared radiation spectrum of polystyrene, showing percent trans-

mission versus radiation frequency, cm–1

Copyright © 1997 by Chemical Education Resources, Inc., P.O. Box 357, 220 S. Railroad, Palmyra, Pennsylvania 17078
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2 TECH 710/Identifying an Unknown Compound by Infrared Spectroscopy

In IR spectroscopy, a simple molecule can produce a very complex

spectrum. An organic chemist takes advantage of this complexity
by comparing the spectrum of an unknown compound with that of a
reference compound. It is unlikely that any two compounds would
produce exactly the same IR spectrum.
Although an IR spectrum is characteristic of the entire molecule, cer-
tain groups of atoms, called functional groups, give rise to particular
absorption bands, or peaks. These absorption bands occur at or near
the same frequency, regardless of the structure of the rest of the mole-
cule. The persistence of these characteristic bands permits chemists to
obtain useful structural information by comparing the absorption bands
for a sample to tables of functional group absorption frequencies.

Studying Vibrations Understanding IR spectroscopy theory requires the study of vibra-

and Energy tion mechanics. In a normal mode of vibration, each atom in a mole-
cule executes a simple harmonic oscillation (vibration) about its equi-
librium position. A ball-and-spring molecular model can be used to
demonstrate the effects of vibrations in stretching and bending
springs, together with the motions of the balls. According to classical
mechanics, the frequency of vibration ν of two balls of total mass m
connected by a spring with a force constant k, is shown in Equation 1.
1
n= ( k / 2 m)1/2 (Eq. 1)
p
The methods of classical mechanics can be used to study the vibra-
tional motions and frequencies of a model containing several balls
(or atoms) of various masses connected by springs (or bonds) with
different force constants. The results of these studies form the basis
for the interpretation of vibrational spectra.
Classical mechanics would indicate that there is a continuum of
vibration levels, and that a molecule may undergo any of numerous
vibrations. Quantum mechanics, however, places restrictions on
microphysical systems. These restrictions limit a molecule to having
discrete energy levels. The difference in energy (∆E) between the
vibrational levels is given by Equation 2, in which h is Planck’s
constant and k is the force constant.
1/2
h k
∆E = hn = khn =   (Eq. 2)
2 p  m
The reduced mass m, defined by Equation 3, is given for a molecule
AB, where MA and MB are the atomic masses of atoms A and B.
MA MB
m= (Eq. 3)
MA + MB
When the frequency of infrared light applied to a compound is ex-
actly the same as the natural vibrational frequency of an interatomic
bond, the molecule absorbs the light and the amplitude of the bond
vibration increases. As indicated in Equation 2, the force constant
for the deformation determines the vibrational frequency. Therefore,
frequency, and thus the frequency of the radiation absorbed, is related
to the rigidity or strength of the bond and the masses of the bonding
atoms. Specifically, the vibrational frequency is higher for stronger
bonds and for lighter atoms.
© 1997 by Chemical Education Resources
 Background Information 3

Two types of vibrations, stretching and bending, are responsible

for most of the important peaks used to identify organic compounds.
For example, Figure 2 shows a few types of vibrations for the CH 2
group. Bending motions require less energy than stretching motions,
so the bending motions absorb at lower frequencies, and, therefore,
have smaller wavenumbers.

Figure 2 Normal modes of CH2 vibra-

tions

The magnitude of infrared absorption bands is proportional to the

change in dipole moment, or separation of charges, that a bond un-
dergoes when it stretches. Thus, the more intense bands in an infrared
spectrum are often produced by C=O and C–O stretching vibrations.
In contrast, the C≡C stretching band for a symmetrical alkyne is al-
most nonexistent because the molecule undergoes no net change in
dipole moment when it stretches.

Interpreting Spectra It is usually not possible to assign specific molecular vibrations to the
majority of bands in an infrared spectrum. However, it is helpful to
divide an IR spectrum into two parts. The 4000 to 1500 cm–1 portion is
useful for identifying various functional groups. Bands in the 1500 to
600 cm–1 portion, called the fingerprint region, are the result of many
types of vibrations that are characteristic of the molecule as a whole.
This complex fingerprint region represents a unique pattern for each
organic compound. The region is useful for comparing the spectrum
of an unknown compound with the spectra of known compounds for
identification purposes.
Two examples, 2-methyl-2-propanol and 2-butanone, illustrate the
application of IR spectroscopy to identify organic functional groups.
In the IR spectrum of 2-methyl-2-propanol, shown in Figure 3(a) on
the next page, the intense band from 3500 to 3100 cm–1 can be attrib-
uted to the OH group. Specifically, this band illustrates the stretching
frequency of the O–H bond. The stretching frequency of a C–H bond
is near 2900 cm–1, and the stretching frequency of a C–O bond is near
1150 cm–1.
The stretching frequency of the C=O group in 2-butanone results in
an intense band near 1700 cm–1, as shown in Figure 3(b) on the next
page. Clearly, the fingerprint regions in Figures 3(a) and 3(b) are dif-
ferent, reflecting the different structures of the two compounds.
Table 1 gives a correlation of the more common structural units and
their characteristic vibrational frequencies.

Measuring IR Spectra Currently, there are two types of instruments used to record IR spectra:
dispersive double-beam and Fourier transformed (FT) spectrophotom-
eters. In a double-beam spectrophotometer, the IR radiation is emitted
into a monochromator, which separates the wavelengths of light. The
monochromatic radiation goes into a beam splitter composed of a mirror
© 1997 by Chemical Education Resources
4 TECH 710/Identifying an Unknown Compound by Infrared Spectroscopy

Table 1 Absorption frequencies of some common bonds (shown in bold type)

bond type of compound frequency

–C–H (stretch) alkanes 2800–3000

=C–H (stretch) alkenes, aromatics 3000–3100

≡C–H (stretch) alkynes 3300
–O– H (stretch) alcohols, phenols 3600–3650 (free)
3200–3500 (H-bonded) (broad)
–O– H (stretch) carboxylic acids 2500–3300
–N–H (stretch) amines 3300–3500 (doublet for NH2)
O
–C–H (stretch) aldehydes 2720 and 2820
–C=C– (stretch) alkenes 1600–1680
–C=C– (stretch) aromatics 1500–1600
–C C–H (stretch) alkynes 2100–2270
O
–C– (stretch) aldehyde, ketones, 1680–1740
carboxylic acids
–C N (stretch) nitriles 2220–2260
C–N (stretch) amines 1180–1360
–C–H (bending) alkanes 1375 (methyl)
–C–H (bending) alkanes 1460 (methyl and methylene)
–C– H (bending) alkanes 1370 and 1385 (isopropyl split)
–C–H (bending) R–CH=CH2 1000–960 and 940–900
–C–H (bending) R2C=CH2 915–870
–C–H (bending) cis RCH=CHR 790–650
–C–H (bending) trans RCH=CHR 990–940
–C– H (out of plane mono subst. benzenes 770–730 and 710–690
bending)
–C– H (out of plane o- subst. benzenes 770-735
bending)
–C– H (out of plane m- subst. benzenes 810–750 and 710–690
bending)
–C– H (out of plane p- subst. benzenes 860–800
bending)
–C– O (stretch) primary alcohols 1050
–C– O (stretch) secondary alcohols 1100
–C– O (stretch) tertiary alcohols 1150
–C– O (stretch) phenols 1200
© 1997 by Chemical Education Resources
 Background Information 5

Figure 3 The IR spectra of (a) 2-methyl-2-propanol and (b) 2-butanone

and a prism. The beam splitter allows equivalent beams of relatively

narrow wavelength range to pass simultaneously through a sample
and a reference cell. Another prism and mirror system focuses the
emergent light into a rotating-sector mirror. An electronic bridge sys-
tem detects the radiant energy transmitted by the sample and the ref-
erence as a voltage difference. A recording of the percent transmission
of the IR radiation through the sample with varying wavelengths pro-
duces an IR spectrum.
The FT-IR method splits the electromagnetic radiation into two
beams. One beam travels over a longer path inside the spectro-
photometer than the other beam. A recombination of the two beams cre-
ates an interference pattern or interferogram. Fourier transformation, a
computerized mathematical manipulation of data from the
interferogram, converts the interferogram into the usual IR spectrum.
The instrument does not use a monochromator. The radiation from the
entire IR spectrum passes through the sample simultaneously, saving
much time. FT-IR instruments can have very high resolution (<0.001
cm–1). Moreover, because the data undergo analog-to-digital conversion,
FT-IR data can be easily manipulated. Combinations of results of several
scans average out random artifacts, allowing excellent spectra from very
small samples.
© 1997 by Chemical Education Resources
6 TECH 710/Identifying an Unknown Compound by Infrared Spectroscopy

Preparing IR Samples Infrared spectra of liquid samples are prepared by placing neat, or
pure, undiluted samples between two salt plates. Glass is opaque to
IR radiation and cannot be used. Instead, the sample is prepared us-
ing potassium bromide (KBr), sodium chloride (NaCl), or silver chlo-
ride (AgCl) plates, which are transparent to IR radiation.
A solid sample can be mixed with solid KBr. The mixture is then
pressed into a very thin pellet. Alternatively, a solid sample can be
dissolved in a solvent such as dichloromethane, trichloromethane,
tetrachloromethane, or carbon disulfide. In a third method, solid sam-
ples may be ground into a very fine powder and mixed with Nujol min-
eral oil to form a mixture called a Nujol® mull. Some solvents do absorb
IR at certain frequencies, so those parts of a spectrum are not useable.
Typically, neat samples for liquids and KBr pellets for solids are
used because neither method produces extraneous absorptions to
obscure the spectra. Consequently, these methods have been chosen
for this experiment.

Equipment
35-mm o. d. agate mortar and pestle 2 KBr, NaCl, or AgCl plates,
gloves with a sample holder
IR spectrophotometer, either lint-free tissues
double-beam or FT microspatula
KBr hand press, with a die set and 6 Pasteur pipets, with latex bulb
an appropriate KBr pellet holder

Reagents and Properties

substance quantity molar mass (g/mol) bp (°C) mp (°C)
p-anisaldehyde* 0.3 mL 136.15 248
benzoic acid 2 mg 122.12 122–123
ethanol* 20 mL
potassium bromide 100 mg
unknown* 0.3 mL
*in a capped vial containing 3Å molecular sieves

Preview
• Prepare a KBr pellet of benzoic acid
• Obtain an IR spectrum of benzoic acid
• Prepare a sample of p-anisaldehyde using either KBr, NaCl, or
AgCl plates
• Obtain a spectrum of p-anisaldehyde
• Prepare a neat IR sample of a liquid unknown
• Obtain an IR spectrum of the liquid unknown
• Deduce the structure of the unknown

PROCEDURE Chemical Alert

p-anisaldehyde—irritant
benzoic acid—irritant
ethanol—flammable and toxic
potassium bromide—irritant and hygroscopic
unknowns—flammable, toxic, corrosive, irritant, and hygroscopic
© 1997 by Chemical Education Resources
 Procedure 7

Caution: Wear departmentally approved safety goggles at all

times while in the chemistry laboratory.

1. Preparing a KBr Pellet Caution: Benzoic acid and KBr are irritants. Prevent eye, skin,
Sample of Benzoic Acid and clothing contact. Avoid inhaling dust and ingesting these
compounds.

View the demonstration amounts of benzoic acid and KBr in the vials
NOTE 1: When preparing IR samples, provided by your laboratory instructor. [NOTE 1] Place approximately
keep everything dry. Do not expose sam- 2 mg of benzoic acid and about 100 mg of dry KBr into a small agate
ples to water. Always keep the bottle
containing the dry, spectroscopic grade mortar. Grind the mixture into a fine powder with a pestle.
KBr in an oven set at 100 °C. When you Place the lower anvil of the hand press with the shorter die pin on a
take the bottle from the oven, cap the clear area of the bench. Attach the collar to the anvil, oriented as
bottle and place it in a desiccator to cool.
shown in Figure 4. [NOTE 2]
NOTE 2: A different type of pellet Use a microspatula to quickly transfer about 75% of the finely
press may be employed in your labora- ground sample into the collar. Place the upper anvil with the longer
tory. If so, your laboratory instructor
will provide instructions for its use. die pin over the collar so that this die pin comes in contact with the
sample, as shown in Figure 4. Place the die set on the hand press
plunger while holding the press in the vertical position.
Slowly compress the sample by pulling down the lever and hold-
ing it for 20–30 s. Release the lever and remove the die set from the
press. Carefully separate the upper and lower anvils. Leave the KBr
pellet in the collar.
Place the collar containing the KBr pellet onto a sample holder. Use the
procedure described in either Part 3A or Part 3B to obtain an IR spectrum.
At the end of the experiment, use a microspatula to remove the KBr
pellet from the collar. Place the pellet material into the container
labeled “Recovered KBr Pellets”, provided by your laboratory
instructor. Wash the metal apparatus carefully with water and dry the
apparatus in the oven.

Figure 4 A KBr hand press with a die

set

© 1997 by Chemical Education Resources

8 TECH 710/Identifying an Unknown Compound by Infrared Spectroscopy

2. Preparing a Sample of A. Using KBr or NaCl Plates

p-Anisaldehyde
Caution: p-Anisaldehyde is an irritant. Prevent eye, skin, and
[NOTE 3]
clothing contact. Avoid inhaling fumes and ingesting the
compound. Use p-anisaldehyde in a fume hood.
NOTE 3: Your laboratory instructor Obtain a sample of p-anisaldehyde from your laboratory instructor.
will tell you which of the following Keep the container capped except for the very brief time necessary to
procedures to use.
remove a sample. To prevent stirring up the molecular sieves, do not
agitate the container prior to use.
NOTE 4: KBr and NaCl plates are frag- Remove a pair of KBr or NaCl plates from the desiccator. [NOTE 4]
ile and hygroscopic. Do not use water to Use a Pasteur pipet to place half a drop of p-anisaldehyde in the center
wipe the plates. Even moisture from your
fingers will attack the plates. Use of one of the plates. Gently press the plates together to remove any air
gloves and only handle the plates by bubbles.
the edges. Place the plates on the sample holder, as shown in Figure 5. Place
the metal cover over the top of the plates and replace all nuts. Gently
tighten the nuts to apply an even pressure to the top plate.
Use the procedure described in either Part 3A or Part 3B to obtain
an IR spectrum.
At the end of the experiment, remove the sample holder from the
sample compartment and unscrew the nuts. Remove the plates and
separate them.
Caution: Absolute ethanol is flammable and toxic. Do not use
near flames or other heat sources. Avoid inhaling fumes and
ingesting the compound.
Use a Pasteur pipet to rinse the plates with dry absolute ethanol.
Then dry the plates with absorbent, lint-free tissues. Place the used
tissues into a container labeled “Used Tissues”, provided by your
laboratory instructor. Return the plates to the desiccator, and return
Figure 5 IR salt plates and holder
the remaining p-anisaldehyde to your laboratory instructor.

B. Using AgCl Plates

Caution: p-Anisaldehyde is an irritant. Prevent eye, skin, and
clothing contact. Avoid inhaling fumes and ingesting the
compound. Use p-anisaldehyde in a fume hood.
NOTE 5: AgCl plates are fragile. Use Obtain two AgCl plates from the desiccator. [NOTE 5] Put half a drop
gloves when handling AgCl plates. of p-anisaldehyde between the plates and gently press them together
Minimize exposure of the plates to light
because light will darken them. to remove any air bubbles.
Assemble the AgCl cell as shown in Figure 6. Place an O-ring in the
cell body and place the AgCl plates on top of the ring. Put the second
O-ring on top of the AgCl plates and then put on the nut. Gently
tighten the nut by hand. Do not overtighten because the AgCl plates
may distort or crack.
Place the fully assembled cell on a sample holder aligning the
assembly so that light can pass through it. Use the procedure de-
scribed in either Part 3A or Part 3B to obtain an IR spectrum.
At the end of the experiment, remove the sample holder from the
sample compartment and unscrew the nut. Remove the AgCl plates
and separate them.
Caution: Absolute ethanol is flammable and toxic. Do not use
near flames or other heat sources. Avoid inhaling fumes and
ingesting the compound.
© 1997 by Chemical Education Resources
 Procedure 9

Figure 6 AgCl cell assembly

Use a Pasteur pipet to rinse the plates with dry absolute ethanol.
Then dry the plates with absorbent, lint-free tissues. Place the used
tissues into a container labeled “Used Tissues”, provided by your
laboratory instructor. Return the KBr plates to the desiccator, and
return the remaining p-anisaldehyde to your laboratory instructor.

3. Obtaining a Spectrum A. Using an FT-IR Spectrophotometer

[NOTE 3]
Make certain that there is nothing in the sample compartment of the
IR spectrophotometer. Use the instructions provided by your labora-
tory instructor to operate the spectrophotometer. Obtain a back-
ground spectrum.
Place the sample holder in the sample compartment and start the
data acquisition. Adjust the output to maximize the display in both
the x and y axes.
Place a new sheet of chart paper on the plotter. Plot the spectrum.
Check with your laboratory instructor to see if the spectrum that
you have recorded is acceptable. If the spectrum is not acceptable,
prepare a new sample and repeat the procedure until you obtain an
acceptable spectrum.

B. Using a Double-Beam IR Spectrophotometer

Place the sample holder in the sample compartment. Use the in-
structions provided by your laboratory instructor to operate the
spectrophotometer.
Place a reference beam attenuator into the reference compartment
to equalize the amount of energy in the two beams. Set the starting
wavenumber to 4000 cm–1 and adjust the attenuator to obtain a read-
ing between 80 and 90% transmission on the chart paper.
Start the scan, watching the pen. If the baseline moves up and the pen
is no longer on the chart, stop the scan and adjust the attenuator to lower
the percent transmission so that the pen is again on the chart. Restart the
scan.
Check with your laboratory instructor to see if the spectrum that
you have recorded is acceptable. If the spectrum is not acceptable,
prepare a new sample and repeat the procedure until you obtain an
acceptable spectrum.
© 1997 by Chemical Education Resources
10 TECH 710/Identifying an Unknown Compound by Infrared Spectroscopy

4. Preparing a Sample of an Caution: Consider the unknown compounds used in this experi-
Unknown Compound Using ment to be flammable, toxic, corrosive, and irritating. Do not
KBr, NaCl, or KBr Plates use them near flames or other heat sources. Prevent eye, skin,
and clothing contact. Avoid inhaling the vapors and ingesting
the compounds. If you spill any unknown, notify your labora-
tory instructor immediately.

Obtain an unknown compound from your laboratory instructor and

record its identification code. Keep the container capped except for
the very brief time necessary to remove a sample. To prevent stirring
up the molecular sieves, do not agitate the container prior to use.
Follow the procedure you used in Part 2A or Part 2B for p-anisal-
dehyde, except use your unknown. Obtain a spectrum for your unknown.
Record the major bands in your spectrum. If you are using an FT-IR
that has a library of standard spectra, compare your spectrum with
the library spectra. Otherwise, compare your spectrum to those in a
list of standard IR spectra provided by your laboratory instructor. Re-
port the identity of your unknown.

5. Cleaning Up Use the labeled collection containers provided by your laboratory

instructor. Place Pasteur pipets in the container labeled “Used Pasteur
Pipets”. Place any tissues used in cleaning KBr or AgCl plates in the
“Used Tissues” container. Do not place tissues in trash. Place used KBr
pellets in the container labeled “Recovered KBr Pellets”.

Caution: Wash your hands thoroughly with soap or detergent

before leaving the laboratory.

Post-Laboratory Questions 1. (a) Clearly label the IR spectrum of benzoic acid with your name,
laboratory section, the date, and “benzoic acid”. Assign the follow-
ing peaks:
broad 3300–2500 cm–1
3020 cm–1
1682 cm–1
(b) Clearly label the IR spectrum of p-anisaldehyde with your
name, laboratory section, the date, and “p-anisaldehyde”. Assign
the following peaks:
3074, 3008 cm–1
2965 cm–1
2834, 2736 cm–1
1695 cm–1
2. For the unknown, make a list of the major peaks in its spectrum and
give your interpretation for each of them.
3. Examine the standard IR spectra of the unknowns that your labora-
tory instructor made available to you to identify your sample.
Examine the spectra to see if you can match the fingerprint region
to that of the spectrum of your unknown. Clearly label the spec-
trum with your name and the unknown identification code.
4. Write the name and structural formula of your unknown.
© 1997 by Chemical Education Resources
 Pre-Laboratory Assignment 11

NAME SECTION DATE

TECH 710/Identifying an Unknown Compound by Infrared Spectroscopy

Pre-Laboratory Assignment

1. Briefly explain the following procedures and safety precautions you should
take in this experiment.
(a) Why are AgCl plates protected from exposure to the light?

(b) Why is it essential that you not handle KBr or NaCl plates with your
bare hands?

(c) What are the hazards of the unknowns?

(d) What will you do if some of an unknown is spilled?

2. Give the frequency ranges (in cm–1) for each of the following:
(a) O–H stretch

(b) carbonyl stretch of an aldehyde

(c) –C–H stretch

(d) =C–H stretch

3. How could you distinguish between cyclohexane and cyclohexene using IR?

© 1997 by Chemical Education Resources

12 TECH 710/Identifying an Unknown Compound by Infrared Spectroscopy

4. Describe the procedure for preparing a liquid sample for infrared

examination.

5. Although IR spectra of solids are most commonly run as KBr pellets, they
are sometimes run as solutions. Even though most organic compounds
are more soluble in acetone than they are in tetrachloromethane (CCl4),
acetone is almost never used as a solvent in running IR spectra of solids.
However, CCl4 is very acceptable provided that the solid will dissolve in
it. Briefly explain why CCl4 is the preferred solvent.

6. If you examine the IR spectrum of an unknown, how could you tell if it is

an alcohol?

ISBN 0-87540-710-2
© 1997 by Chemical Education Resources