Reagent Chemicals

76 Infrared Spectroscopy

Resolution, R, can be specified to ensure separation of closely eluting components or to establish the general separation efficiency of the system. R can be calculated as follows:

R=

. (t 2 t 1)
1178
W(h /2)1 + W(h /2)2

where t2 and t1 are the retention times of the two component peaks, and W(h/2)1
and W(h/2)2 are the half-height widths of the two peaks in units of time. An R value
of 1.0 means that the resolution is 98% complete. This condition is sufficient, in
most cases, for peak area calculations. An R value of 1.5 or greater represents
baseline, or complete, separation of the peaks.

INFRARED SPECTROSCOPY
Infrared (IR) spectroscopy is an absorption method widely used in both qualitative and quantitative analysis. The infrared region of the spectrum includes wave
numbers ranging from about 12,800 to 10 cm1. The IR range can be separated
into three regions: the near-infrared (12,8004,000 cm1), the mid-infrared
(4,000200 cm1), and the far-infrared (20010 cm1). Most analytical applications fall in the mid-infrared region of the spectrum. IR spectroscopy has primarily been used to assist in identification of organic compounds. The IR spectrum
of an organic compound is a unique physical property and can be used to identify
unknowns by interpretation of characteristic absorbances and comparison to
spectral libraries. IR spectroscopy is also employed in quantitative techniques.
Because of its sensitivity and selectivity, IR spectroscopy can be used to quantitate
analytes in complex mixtures. Quantitative analysis is used extensively in detection of industrial pollutants in the environment. The near-infrared region has
been used for quantitative applications, such as determination of water in glycerol
and determination of aromatic amines in complex mixtures. The IR technique is
discussed here primarily for application in identification of organic compounds
and will focus on the mid-infrared region. Instrumental operating procedures are
not given because they will vary depending on instrument design. A brief discussion of the theory will be followed by a discussion of instrumentation, sample
handling techniques, and qualitative analysis.

General Background
Unlike UV and visible spectroscopy, which uses larger energy absorbances from
electronic transitions, IR spectroscopy relies on the much smaller energy absorbances that occur between various vibrational and rotational states. When molecular vibrational or rotational events occur that cause a net dipole moment, IR
absorption can occur. Molecular vibrations can be classified as either stretching or

Ninth Edition

ACS Specifications

Official from January 1, 2000

2000 American Chemical Society

Infrared Spectroscopy

77

bending. Stretching is a result of continuous changing distances in a bond between

two atoms. Bending refers to a change in the angle between two bonds. Bending
motions include scissoring, rocking, wagging, and twisting. The various types of
vibrations and rotations absorb at different frequencies within the infrared region,
thus resulting in unique spectral properties for different molecular species.

Instrumentation
Basic instrumentation for IR spectroscopy includes a radiation source, wavelength selector, sample container, detector, and signal processor. Continuous
sources are used in the mid-infrared region and include the incandescent wire
source, the Nernst glower, and the Globar. The Nernst glower is hotter and
brighter than the incandescent wire. The Globar is a rod of silicon carbide that is
electrically heated and that typically requires a water cooling system. The Globar
provides greater output than the Nernst glower in the region below 5 mm. Otherwise, their spectral energies are similar. Newer technologies have also employed
various lasers as sources for infrared applications.
The three general types of infrared detectors are thermal, pyroelectric, and
photoconducting detectors. Thermal detectors measure very minute temperature
changes as a method of detection. They operate over a wide range of wavelengths
and can be operated at room temperature. The main disadvantages of thermal
detectors are slow response and less sensitivity when compared to other detectors.
Pyroelectric detectors are specialized thermal detectors that provide faster
response and more sensitivity. Photoconducting detectors rely on interactions
between incident photons and a semiconductor material. These detectors also
provide increased sensitivity and faster response.
The two main types of instruments used for qualitative analysis are dispersive-grating spectrophotometers and multiplex instruments that employ Fourier
transform. Wavelength selection in dispersive-grating instruments is accomplished using filters, prisms, or reflection gratings. Traditional instruments have
consisted of a filter-grating or prism-grating system that covers the range between
4000650 cm1. The most common is a double-beam instrument that uses reflection grating for dispersing radiation.
Fourier transform has been applied to infrared spectroscopy and is now a
very common technique. Fourier transform infrared (FTIR) spectroscopy offers
enhanced sensitivity compared to dispersive IR spectroscopy. Signal-to-noise
ratios are often improved by an order of magnitude. Most commercial FTIR
instruments are based on the Michelson interferometer and require a computer
interface to perform the Fourier transform and process data. Using an interferometer provides an increase in resolution and results in accurate and reproducible frequency determinations. This offers an advantage when using background
subtraction techniques. Most Fourier transform instruments are now benchtop
size and relatively easy to maintain. These instruments are largely replacing traditional dispersive instruments.

Reagent Chemicals
Ninth Edition

ACS Specifications

Official from January 1, 2000

2000 American Chemical Society

78 Infrared Spectroscopy

Sample Handling
Sample containers and handling can present a challenge in the infrared region.
Materials used to produce cuvettes are not transparent and cannot be used. Cells
prepared from alkali halides such as sodium chloride are widely used due to their
transparent properties in the infrared region. A common problem with sodium
chloride cells is that they absorb moisture and become fogged. Polishing is
required to restore the cells to a more transparent state.
Liquids may be analyzed in their neat form by placing a small amount of
sample on a sodium chloride plate and then placing a second plate on top to form
a sample film. The plates are then placed in an appropriate holder in the sample
compartment of the instrument. This technique provides adequate spectra for
qualitative use.
Solutions of liquid or solid materials can also be analyzed by IR spectroscopy.
Solvents should be chosen that do not have absorbances in the region of interest.
Unfortunately, no solvent is completely transparent in the mid-infrared region.
With double-beam instruments, a reference cell containing blank solvent can be
employed. Common moderate absorbances will not be observed. Solvent transmission should always be above 10% when using a solvent reference cell. Influences of solvent on the absorbance of the solute should be considered. For example, hydrogen bonding of alcohols or amines with the solvent may affect
characteristic vibrational frequency of the functional group. When practical, it is
desirable to analyze neat materials for qualitative analysis.
A method commonly used for analysis of neat solid samples is the mull
technique. The technique consists of grinding the material into a fine powder
and then dispersing it into a liquid or solid matrix to form a mull. Liquid mulls
have been formed by combining the powdered analyte with Nujol (a heavy
hydrocarbon oil). The liquid mull is analyzed between salt plates as described
above. The disadvantage of Nujol is that hydrocarbon bands may interfere with
analyte absorbances. A second method of forming a mull involves grinding the
powdered analyte with dry potassium bromide and forming a disk. The ratio of
analyte to potassium bromide is usually about 1:100. The materials are ground
together using a mortar and pestle or a small ball mill. The mixture is then
pressed in a die at 10,00015,000 psi to form a small transparent disk and analyzed. Care must be taken when preparing the disk to protect it from moisture.
It is very common to see absorbances for moisture when using potassium bromide disks.
Another technique for handling samples is the use of disposable sample
cards. These commercially available cards contain an IR-transparent material on
which a neat liquid or solution can be placed for analysis. An analyte can be dissolved in a volatile solvent, placed on the card, and the solvent evaporated to form
a thin coating of sample, which can then be analyzed. This technique may be used
when a limited amount of analyte material is available.

Reagent Chemicals
Ninth Edition

ACS Specifications

Official from January 1, 2000

2000 American Chemical Society

Infrared Spectroscopy

79

Qualitative Analysis
The most widely used application of IR spectroscopy is for qualitative analysis of
organic compounds. Compounds have unique spectra that depend on molecular
attributes. A common method of interpreting IR spectra is to consider two
regions: the functional group frequency region (36001200 cm1) and the fingerprint region (1200600 cm1). A combination of interpreting the functional
group region and comparing the fingerprint region with those in spectral libraries provides, in many cases, sufficient evidence to positively identify a compound.
The functional group region provides evidence of functional groups in a
molecule based on the absorbance frequency. Tables are available in standard
spectra manuals that provide ranges of absorbances for specific functional
groups. Common groups with characteristic absorbances include aldehydes,
ketones, esters, alkenes, alkynes, alcohols, amines, amides, carboxylic acids, nitro
groups, and nitriles. While most functional groups fall in the 36001200 cm1
range, some can also fall in the fingerprint region. For example, CO bonds can
be around 1000 cm1, and CCl absorbances are typically found in the range of
600800 cm1.
The fingerprint region is often unique to the analyte. In this region, very
small differences in structure can lead to differences in absorbances. Most single
bonds absorb in this area, and differences in skeletal structure of molecules will
result in frequency and intensity differences. The fingerprint region is most effectively used by comparison to existing spectra. Many commercial suppliers if IR
instrumentation also offer searchable electronic spectral libraries. Computerbased search systems offer a very rapid method of comparing unknown samples
to known spectra. The systems usually yield a list of possible compounds ranked
in order of best fit of the unknown spectrum to the library spectrum.
Procedure
IR spectroscopy is used in selected standard-grade reference materials specifications as a technique for identity confirmation. The criteria under which standardgrade reference materials pass identity tests are based on observation of characteristic absorbances and by comparison to a spectral library, if one is available. Each
entry to be analyzed by IR spectroscopy will contain a minimum of three absorbances that must be present to pass the test. In addition, NIST should be used as a
source to obtain comparison spectra. A standard-grade reference material will
pass the comparison test if all absorbances found in the NIST spectrum are
present in the test spectrum. The NIST spectral library contains many of the standard-grade reference materials listed in this book, and spectral comparisons
should be made if NIST spectra are available. NIST spectra are available by contacting NIST or by accessing NISTs Web site, http://webbook.nist.gov/chemistry/.

Reagent Chemicals
Ninth Edition

ACS Specifications

Official from January 1, 2000

2000 American Chemical Society