2.
Instrumentation
Optical spectroscopy is based on
Absorption
Fluorescence
Phosphorescence
Emission
Chemiluminescence
The instrument for each method is different, but
basic components are the same.
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Components of optical Instruments
Stable source of radiant energy.
Device that isolates wavelengths of light for
measurement.
A transparent container for holding the sample.
A radiation detector, which converts radiant energy to
electrical signal.
A signal processor and readout.
Light Wavelength Sample Radiation Signal
Source selector Holder detector processor
NOTE: The wavelength selector can also come after the sample
holder. 2
Absorption measurement
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Fluorescence measurement
Two wavelength
selectors are needed
in order to isolate the
excitation and
emission wavelengths
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Emission measurement
A source of thermal
energy, such as a flame
or plasma, produces an
analyte vapor that emit
radiation isolated by a
wavelength selector
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Light sources: produce beam with sufficient power
Continuum sources – emit radiation that changes
intensity only slowly as a function of
H2 or D2 lamps give light in the UV region.
Tungsten filament lamp is very commonly used for visible
light.
Inert solids heated to 1500-2000K make common IR
sources.
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Light sources: produce beam with sufficient power
Line Sources – emit limited number of lines or band of radiation
that is used in AAS, atomic and molecular fluorescence
Mercury and sodium vapor lamps provide a few sharp lines in
the UV and Vis regions.
Hollow cathode lamps
Laser Sources (Light Amplification by Stimulate Emission of
Radiation)
– Lasers: have high intensities, narrow bandwidths, and
coherent output.
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Wavelength Selectors
Radiation that consists of
narrow, continuous group of
wavelengths is required for
most spectroscopic analyses.
Narrow bandwidth enhances
the sensitivity of absorbance
measurement.
Having a narrow band ensures
a linear relationship between
optical signal and
concentration.
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Monochromators
Spectroscopic methods: to vary of
radiation continuously (scanning).
Entrance slit: provides a optical
image.
A collimating lens or mirror:
produces a parallel beam of
radiation.
A prism or grating: disperses the
radiation into its component
wavelengths.
Focusing element: reforms the
image of the entrance slit.
An exit slit: isolates the desired
spectral band.
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Monochromator slits
The entrance slit serves as a radiation source.
The exit slit serves the purpose of selecting the wavelength.
Moving the exit slit, defines the angle (and consequently
the wavelength) of the reflected (or bent) light.
• Most monochromators have variable slits.
– Need a minimal slit width to resolve narrow absorption or
emission bands.
– Wider slits are needed to let more light in and produce
enough radiative power to be measurable.
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Transparent sample containers
Sample containers need to be transparent in the
spectral region of interest.
Quartz or fused silica is used in the UV region.
Silicate glass is used between 350 and 2000 nm.
Crystalline salts (NaCl, KBr) are used for cell windows
in the IR region.
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Transducers (detectors)
Earliest detector: eye, photographic plate or
films
Convert radiation energy into an electrical
signal
An ideal transducer should have
– High sensitivity
– High S/N
– Fast response
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Radiation Transducers
• Photomultiplier tubes: are very
sensitive to low radiant power.
Light strikes a photoemission
cathode.
The ejected electrons strike a
dynode causing the emission
of several additional
electrons.
The process is repeated many
times to get a large number of
ejected electrons per photon
of light.
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Thermal transducers
Radiation transducers: don’t work well in the IR
because the IR photons lack the energy to cause
photoemission of electrons.
Thermal transducers: work by absorbing the radiation
to give a temperature increase which is measured.
Thermistors
Constructed of a material that exhibits a large change in
resistance as a function of temperature.
Pyroelectric transducers
A pyroelectric crystal generates an electrical charge when
heated. 14
Signal processors
The signal processor is an electronic device that
amplifies the electrical signal from the transducer.
It may alter the signal from dc to ac or the reverse.
May perform mathematical operations on signals
such as differentiation, integration, or conversion to
logarithms.
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Types of optical Instruments
Colorimeter
An instrument for absorption measurements in which the
human eye serves as the detector using one or more color-
comparison standards.
Spectrometer
An instrument that provides information about the intensity
of radiation as a function of wavelength or frequency.
Spectrophotometer
A spectrometer that is equipped with one or more exit slits
and photoelectric transducers that permit the
determination of the ratio of the power of two beams as a
function of wavelength.
A spectrophotometer for fluorescence analysis is called a
spectrofluorometer. 16
Single-beam versus double-beam spectrophotometer
Single-beam spectrophotometer
Equipped with a fixed-
wavelength monochromator
The wavelength is adjusted
manually
Limitation:
Not handy and time-consuming
Accuracy depends on the
stability of light source
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Double-beam spectrophotometer
A chopper controls the
radiation’s path, alternating
it between the sample, the
blank and a shutter.
The signal processor uses
the chopper’s known speed
of rotation to resolve the
signal reaching the
detector into transmission
of the blank, P0, and the
sample, PT
An opaque surface as a
shutter: continuously
adjust 0% T.
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