ULTRAVIOLET-VISIBLE SPECTROPHOTOMETERS

1. SINGLE-BEAM SPECTROPHOTOMETER

Sample= analyte + solvent; Blank= solvent alone

• The simplest spectrophotometer is a single-beam instrument equipped with a fixed

wavelength monochromator, the block diagram for which is shown above.
• Single optical path between the source and detector.
• Calibrated to 0% T while using a shutter to block the source radiation from the detector.
• Shutter removed, the instrument is calibrated to 100% T using an appropriate blank.
• The blank is then replaced with the sample, and its transmittance is measured. Since the
source’s incident power and the sensitivity of the detector vary with wavelength, the
photometer must be recalibrated whenever the filter is changed.
• The limitations of fixed-wavelength, single-beam spectrophotometers are
minimized by using the double-beam spectrophotometer.

2. (a) DOUBLE-BEAM SPECTROPHOTOMETER (IN-SPACE)

• The figure below shows a double-beam-in-space instrument in which two beams are
formed by a V-shaped mirror called a beam-splitter.
• One beam passes through the reference solution to a photodetector, and the second
simultaneously passes through the sample to a second, matched photodetector.

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 • The two outputs are amplified, and their ratio, or the logarithm of their ratio, is obtained
electronically or computed and displayed on the output device.
• NOTE: Because the original beam is split into two equal parts, the energy or intensity is
reduced by half. This is why space sharing is infrequently used.

2. (b) DOUBLE-BEAM SPECTROPHOTOMETER (IN-TIME)

• In this design, the beam goes to the blank compartment at one time and at the other time
to the sample compartment.
• The beams are separated in time by a rotating sector mirror (chopper) that directs the
entire beam through the reference cell and then through the sample cell. The pulses of
radiation are then recombined by another mirror that transmits the reference beam and
reflects the sample beam to the detector.
• The instrument substracts the blank reading electronically so that the only signal of
importance is that of the sample alone.
• The double-beam-in-time approach is generally preferred over the double-beam-in-space
because of the difficulty in matching two detectors.

Advantages of double beam instruments:

1. compensate for slow fluctuations in the radiation from the source

2. compensate for wide variations of source intensity with wavelength
3. Detector response is more uniform than in single beam designs
4. Amplifier gain is also more uniform than in single beam instruments.
5. double-beam design is well suited for continuous recording of absorption spectra.

Disadvantages: high cost, complexity and heavy maintenance of the double beam
spectrophotometer.

• QUANTITATIVE ANALYSIS FOR A SINGLE ANALYTE AND MIXTURES

o Refer to the notes and worked examples in the Beer-Lambert Law section

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Note: In ultraviolet/visible instruments, cells are always located between the monochromator and
the detector in order to avoid photochemical decomposition, which may occur if samples are
exposed to the full power of an ultraviolet or visible source.

INFRARED SPECTROPHOTOMETERS

Radiation Source
The most common sources of IR radiation for the mid-IR region are Nernst glowers,
Globars, and heated wires and lasers.
• The Nernst filament is fabricated from a binder and oxides of thorium, cerium, zirconium
and yttrium. The disadvantage of Nernst glower is that it is non-conducting at ambient
temperature.
• Globar is a small rod of silicon carbide usually 5 cm in length and 0·5 cm in diameter.
The maximum radiation for the Globar occurs in the 5500-5000 cm-1 region.
• Nichrome wire, carbon arc, rhodium wire and tungsten filament lamp are also used as
light source.
The normal operating temperatures for IR sources are between 1100 and 1500 K. The range of
light put out by mid-IR sources extends into both the NIR and far-IR regions, but intensity is at a
maximum in the mid-IR region from 4000 to 400 cm-1
Monochromators and Interferometers
The radiation emitted by the source covers a wide frequency range. However, the sample absorbs
only at certain characteristic frequencies.
• Monochromators select radiation of any desired frequency from the source and eliminate
that at other frequencies.
• IR spectrometers in current use are FTIR based on a Michelson interferometer.

FOURIER TRANSFORM (FT) SPECTROMETERS

In n Fourier transform infrared spectrometer,

or FT–IR, the monochromator is replaced
with an interferometer to obtain a
spectrum. Fourier transform instruments
contain no dispersing element, and all
wavelengths are detected and measured
simultaneously using a Michelson
interferometer.

Schematic of an interferometer for FTIR spectrometry

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 • The basis of an interferometer - Radiation from an IR source is split into two paths by a
beam splitter, one path going to a fixed position mirror and the other to a moving mirror.
• When the beams are reflected, one is slightly displaced (out of phase) from the other
since it travels a smaller (or greater) distance due to the moving mirror.
• These beams recombine to produce an interference pattern (of all wavelengths in the
beam) before passing through the sample.
• The wavelengths are observed at the sample simultaneously and the interference pattern
changes with time as the mirror is continuously scanned at a linear velocity.
• The result of the absorption of the radiation by the sample is a spectrum in the time
domain called an interferogram (spectrum in the time domain). Fourier transformation
converts it into the frequency domain.

An interferogram obtained from a FT-

IR spectrometer. The plot shows
detector signal output as a function of
time.

Because an FT–IR includes only a single optical path, it is necessary to collect a separate
spectrum to compensate for the absorbance of atmospheric CO2 and H2O vapor. This is done by
collecting a background spectrum without the sample and storing the result in the instrument’s
computer memory. The background spectrum is removed from the sample’s spectrum.

Fourier transform IR spectrum of a thin film of polystyrene. Note that the Fourier transform
takes signal intensity collected as a function of time and produces transmittance as a function of
frequency after subtraction of a background interferogram and proper scaling.

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Software and databases of spectra can be installed and used to process spectral data and to match
measured spectra to known spectra in the database.

Qualitative applications of infrared spectrometry:

1. Identifying functional groups in a molecule

2. Quantitative analysis of mixtures of closely related organic compounds e.g atmospheric
pollutants

Advantages of FT Systems

1. Compared with dispersive systems FT spectrometers produce better S/N ratios. This
results from several factors. FT instruments have fewer optical elements and no slits, so
the intensity of the radiation reaching the detector is much higher than in dispersive
instruments. The increase in signal increases the S/N ratio.
2. All available wavelengths are measured simultaneously, so the time needed to collect all
the data to form a spectrum is drastically reduced. An entire IR spectrum can be collected
in less than 1 s. This makes practical the collection and signal averaging of hundreds of
repetitions of the spectrum measurement. Improvement in S/N from signal averaging
attained.
3. FT spectrometers have high wavelength reproducibility. In an FTIR spectrometer, the
position of the mobile mirror during its motion is continuously calibrated with extreme
precision using highly monochromatic light from a visible range laser. This accurate
position measurement translates into accurate and reproducible analytical wavelength
measurements after Fourier transformation of the interferogram. This accurate position
measurement permits the precise addition of multiple spectra to achieve the multiplex
advantage.