Unit 2
Instruments for optical spectroscopy
The basic components of analytical instruments for absorption spectroscopy, as well as for
emission and fluorescence spectroscopy; are remarkably alike in function and in general
performance requirements whether the instruments are designed for ultraviolet (UV), visible, or
infrared (IR) radiation. Because of the similarities, such instruments are frequently referred to as
optical instruments even though the eye is sensitive only to the visible region.
Instrument Components
Most spectroscopic instruments for use in the UV /visible and IR regions are made up of five
components: 1) a stable source of radiant energy: 2) a wavelength selector that isolates a limited
region of the spectrum for measurement: 3) one or more sample containers: 4) a radiation
detector, which converts radiant energy to a measurable electrical signal: and 5) a signal
processing and readout unit, usually consisting of electronic hardware and, in modern
instruments, a computer.
Optical Materials
The cells, windows, lenses, mirrors, and wavelength-selecting elements in an optical
spectroscopic instrument must transmit radiation in the wavelength region being investigated.
The following figure shows the usable wavelength range for several optical materials that are
used in the UV, visible, and IR regions of the spectrum. Ordinary silicate glass is completely
adequate for use in the visible region and has considerable advantage of low cost. In the UV
region at wavelengths shorter than about 380 nm, glass begins to absorb and fused silica or
quartz must be substituted. Also, in the IR region, glass, quartz, and fused silica all absorb at
wavelengths longer than about 2.5 µm. Hence, optical elements for IR spectrometry are typically
made from halide salts or in some cases, polymeric materials.
Basic Components of Spectroscopic Instrumentation
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Spectroscopic Sources
All forms of spectroscopy require a source of energy. In absorption and scattering spectroscopy
this energy is supplied by photons (electromagnetic radiation). Emission and luminescence
spectroscopy use thermal, radiant (photon), or chemical energy to promote the analyte to a less
stable, higher energy state.
Spectroscopic sources are of two types: continuum sources, which emit radiation that changes in
intensity only slowly as a function of wavelength, and line sources, which emit a limited number
of spectral lines, each of which spans a very limited wavelength range. Sources can also be
classified as continuous sources, which emit radiation continuously with time, or pulsed sources,
which emit radiation in bursts.
Figure: Emission spectrum from a typical continuum source
Figure: Emission spectrum from a typical line source.
Sources of Thermal Energy: The most common sources of thermal energy are flames and
plasmas. Flame sources use the combustion of a fuel and an oxidant such as acetylene and air, to
achieve temperatures of 2000–3400 K. Plasmas, which are hot, ionized gases, provide
temperatures of 6000–10,000 K.
Chemical Sources of Energy: Exothermic reactions also may serve as a source of energy. In
chemiluminescence the analyte is raised to a higher-energy state by means of a chemical
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reaction, emitting characteristic radiation when it returns to a lower-energy state. When the
chemical reaction results from a biological or enzymatic reaction, the emission of radiation is
called bioluminescence. Commercially available “light sticks” and the flash of light from a
firefly are examples of chemiluminescence and bioluminescence, respectively.
Continuum Sources in the Ultraviolet/Visible Region
An ordinary tungsten filament lamp provides a distribution of wavelength from 320 to 2500 nm.
Generally, these lamps are operated at a temperature of around 2900 K which produces useful
radiation from about 350 to 2200 nm.
Tungsten/halogen lamps, also called quartz/halogen lamps, contain a small amount of iodine
within the quartz envelope that houses the filament. Quartz allows the filament to be operated at
a temperature of about 3500 K, which leads to higher intensities and extends the range of the
lamp well into the UV region. The lifetime of a tungsten/halogen lamp is more than double that
of an ordinary tungsten lamp because the life of the latter is limited by sublimation of tungsten
from the filament. In the presence of iodine, the sublimed tungsten reacts to give gaseous WI 2
molecules, which then diffuse back to the hot filament, where they decompose and redeposit as
W atoms. These lamps are finding ever-increasing use in modern spectroscopic instruments
because of their extended wavelength range, greater intensity, and longer life.
Deuterium (and also hydrogen) lamps are most often used to provide continuum radiation in the
UV region. A deuterium lamp consists of a cylindrical tube containing deuterium at low
pressure, with a quartz window from which the radiation exits. The mechanism by which a
continuum is produced by this source involves the formation of an excited molecule D* 2 (or H*2)
by absorption of electrical energy. This species then dissociates to give two hydrogen or
deuterium atoms plus an ultraviolet photon. Both deuterium and hydrogen lamps provide a useful
continuum spectrum in the region from 160 to 375 nm. The deuterium lamp is more widely used
than the hydrogen lamp because the deuterium lamp is more intense.
where Ee is the electrical energy absorbed by the molecule.
In addition to the continuum sources just discussed, line sources are also important for use in the
UV/visible region. Low-pressure mercury arc lamps are very common sources that are used in
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liquid chromatography detectors. The dominant line emitted by these sources is the 253.7-nm Hg
line. Hollow-cathode lamps are also common line sources that are specifically used for atomic
absorption spectroscopy. Lasers have also been used in molecular and atomic spectroscopy, both
for single-wavelength and for scanning applications.
Hollow cathode lamp (HCL)
An electrical potential is applied between the anode and cathode and some of the fill gas atoms
are ionized. The positively charged ions collide with the negatively charged cathode and dislodge
metal atoms in a process called "sputtering." Sputtered metal atoms are further excited to
emission through impact with the fill gas.
Continuum Sources in the Infrared Region
The continuum sources for IR radiation are normally heated inert solids. A Globar source is a
silicon carbide rod; infrared radiation is emitted when the Globar is heated to about 1500°C by
the passage of electricity. A Nernst glower is a cylinder of zirconium and yttrium oxides that
emits IR radiation when heated to a high temperature by an electric Current. Electrically heated
pirals of nichrome wire also serve as inexpensive IR sources.
Continuum Sources
Ar Lamp VAC UV
Xe Lamp VAC UV, UV-VIS
H2 or D2 Lamp UV
Tungsten Lamp UV-Near IR
Nernst Glower UV-VIS-Near IR-IR
Nichrome Wire Near IR-Far IR
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Globar Near IR-Far IR
Line Sources
Hollow Cathode Lamp UV-VIS
Lasers UV-VIS-Near IR
Na and Hg vapor lamps UV-VIS
Electrode less discharge lamps UV-VIS
Wavelength Selectors
Spectroscopic instruments in the UV and visible regions are usually equipped with one or more
devices to restrict the radiation being measured to a narrow band that is absorbed or emitted by
the analyte. Such devices greatly enhance both the selectivity and the sensitivity of an
instrument. In addition, for absorption measurements narrow bands of radiation greatly diminish
the chance of Beer's Law deviations due to polychromatic radiation. Many instruments use a
monochromator or filter to isolate the desired wavelength band so that only the band of interest is
detected and measured.
Radiation Filters
Filters operate by absorbing all but a restricted band of radiation from a continuum source. Two
types of filters are used in spectrometer: Interference filters and Absorption filters.
Interference Filters: They rely on optical interference to provide narrow bands of radiation. It
consists of a transparent dielectric that occupies the space between two semitransparent metallic
films. They are available with transmitter peaks throughout the ultraviolet region and visible
regions and up to about 14m in the infrared.
Interference Wedges: An interference wedge consists of a pair of mirrored, partially transparent
plates separated by a wedge-shaped layer of a dielectric material. They are available for the
visible region, the near-infrared region, and for several parts of the infrared region. They can
serve in place of prisms or gratings in monochromators.
Absorption Filters: They are generally less expensive than interference filters and are widely
used for band selection in the visible region. They function by absorbing certain portions of the
spectrum. The most common type consists of colored glass or of a dye suspended in gelatin and
sandwiched between glass plates. A purple filter, for example, removes the complementary
color green from 500–560 nm. Commercially available absorption filters provide effective
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bandwidths from 30–250 nm. The maximum throughput for the smallest effective bandpasses,
however, may be only 10% of the source’s emission intensity over that range of wavelengths.
The former have the advantage of greater thermal stability. Interference filters are more
expensive than absorption filters, have narrower effective bandwidths, typically 10–20 nm, with
maximum throughputs of at least 40%. One limitation of an absorption or interference filter is
that they do not allow for a continuous selection of wavelength. If measurements need to be
made at two wavelengths, then the filter must be changed in between measurements. A further
limitation is that filters are available for only selected nominal ranges of wavelengths.
Wavelength Selection Using Monochromators: An alternative approach to wavelength
selection, which provides for a continuous variation of wavelength, is the monochromator.
Monochromators generally have a diffraction grating to disperse the radiation into its component
wavelengths. Radiation from the source enters the monochromator through an entrance slit. The
radiation is collected by a collimating mirror, which reflects a parallel beam of radiation to a
diffraction grating. The diffraction grating is an optically reflecting surface with a large number
of parallel grooves. Diffraction by the grating disperses the radiation in space, where a second
mirror focuses the radiation onto a planar surface containing an exit slit. In some
monochromators a prism is used in place of the diffraction grating. Radiation exits the
monochromator and passes to the detector.
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Figure: Typical Monochromator
A polychromatic source of radiation at the entrance slit is converted at the exit slit to a
monochromatic source of finite effective bandwidth. The choice of which wavelength exits the
monochromator is determined by rotating the diffraction grating. A narrower exit slit provides a
smaller effective bandwidth and better resolution, but allows a smaller throughput of radiation.
Monochromators are classified as either fixed-wavelength or scanning. In a fixed-wavelength
monochromator, the wavelength is selected by manually rotating the grating. Normally, a fixed-
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wavelength monochromator is only used for quantitative analyses where measurements are made
at one or two wavelengths. A scanning monochromator includes a drive mechanism that
continuously rotates the grating, allowing successive wavelengths to exit from the
monochromator. Scanning monochromators are used to acquire spectra and, when operated in a
fixed wavelength mode, for quantitative analysis.
Prism Monochromators: Can be used to disperse ultraviolet, visible, and infrared radiation.
1. Dispersing prisms: separation of wavelengths due to differences in index of refraction of
the glass in the prism with each different wavelength. This leads to constructive and
destructive interference. Dispersion is angular (nonlinear). Single order is obtained. The
larger the focal length, the better the dispersion.
2. Reflecting prisms: designed to change direction of propagation of beam, orientation, or
both
3. Polarizing prisms: made of birefringent materials
Grating Monochromators: Can be considered as a set of slits at which diffraction occurs and
destructive/constructive interference occurs that yields a diffraction pattern. Grooves patterns
are now generated by machine. They are manufactured by ruling a piece of glass or metal.
Transmission grating: by directing a polychromatic beam through it, dispersion of ultraviolet,
visible, and infrared radiation can be brought.
Reflection grating- used essentially for the same reason as transmission but is more common
than transmission.
Replica grating: used most in monochromators; manufactured from a master grating. Laying
down a polymer film over it to copy the groove pattern produces them. Replicas are what are
actually used in instruments due to the great difficulty and cost of achieving high quality
gratings.
Sample Containers:
Sample containers are required for all spectroscopic studies except emission spectroscopy. In
common with the optical elements of monochromators, the cells or cuvettes that hold the
samples must be made of material that passes radiation in the spectral region of interest. Quartz
or fused silica is required for work in the ultraviolet re-ion (below 150 nm); both of these
substances are transparent in the visible region and up to about 3µm in the infrared region as
well. Silicate glasses can be employed in the region between 350 and 2000nm. Plastic
containers have also found application in the visible region. Crystalline sodium chloride is the
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most common substance employed for cell windows in the infrared region. Sample containers
must be made of material that is transparent to the spectral region of interest. The best cells have
windows that are perpendicular to the direction of the beam in order to minimize reflection
losses. The most common cell path length for studies in the UV and visible regions is 1 cm;
matched, calibrated cells of this size are available from several commercial sources. Many other
cells with shorter and longer path lengths can be purchased.
Particular care must be taken to duplicate the position of such cells with respect to the beam;
otherwise, variations in path length and reflection losses at the curved surfaces can cause
significant error. The quality of spectroscopic data is critically dependent on the way that cells
are used and maintained. Fingerprints, grease, or other deposits on the walls markedly alter the
transmission characteristics of a cell. Thus, thorough cleaning before and after use is imperative,
and care must be taken to avoid touching the windows after cleaning is complete. Matched cells
should never be dried by heating in an oven or over a flame because this may cause physical
damage or a change in path length. Matched cells should be calibrated against each other
regularly with an absorbing solution.
Detector/Radiation Transducers
A detector is a device that indicates the existence of some physical phenomenon. To obtain
spectroscopic information, the radiant power transmitted, fluoresced, or emitted must be detected
in some manner and converted into a measurable quantity. The human eye is also a detector; it
converts visible radiation into an electrical signal that is passed to the brain via a chain of
neurons in the optic nerve and produces vision. The term transducer is used to indicate the type
of detector that converts quantities, such as light intensity, pH, mass, and temperature, into
electrical signals that can be subsequently amplified, manipulated, and finally converted into
numbers proportional to the magnitude of the original quantity.
Ideally: High sensitivity, High S/N, Constant response over range of wavelengths, Fast response,
Zero output in absence of illumination, Electrical signal directly proportional to radiant power.
Types of Transducers
There are two general types of transducers: one type responds to photons, the other to heat. All
photon detectors are based on the interaction of radiation with a reactive surface either to
produce electrons (photoemission) or to promote electrons to energy states in which they can
conduct electricity (photoconduction). Only UV, visible, and near-IR radiation possess enough
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energy to cause photoemission to occur; thus, photoemissive detectors are limited to wavelengths
shorter than about 2 μm (2000 nm). Photoconductors can be used in the near-, mid-, and far-IR
regions of the spectrum. Generally, we detect IR radiation by measuring the temperature rise of a
blackened material located in the path of the beam or by measuring the increase in electrical
conductivity of a photoconducting material when it absorbs IR radiation.
Photon Detectors: Widely used types of photon detectors include phototubes, photomultiplier
tubes, silicon photodiodes, and photodiode arrays.
Phototubes and Photomultiplier Tubes: The response of a phototube or a photomultiplier tube
is based on the photoelectric effect. A phototube consists of a semicylindrical photocathode and
a wire anode sealed inside an evacuated transparent glass or quartz envelope. The concave
surface of the cathode supports a layer of photoemissive material, such as an alkali metal or a
metal oxide, which emits electrons when irradiated with light of the appropriate energy. When a
voltage is applied across the electrodes, the emitted photoelectrons are attracted to the positively
charged wire anode. A photocurrent then results that is easily amplified and measured. The
number of photoelectrons ejected from the photocathode per unit time is directly proportional to
the radiant power of the beam striking the surface. With an applied voltage of about 90 V or
more, all these photoelectrons are collected at the anode to give a photocurrent that is also
proportional to the radiant power of the beam.
Figure: A phototube and accompanying circuit. The photocurrent induced by the radiation causes
voltage across the measuring resistor: this voltage is then amplified and measured.
The photomultiplier tube (PMT) is similar in construction to the phototube but is significantly
more sensitive. Its photocathode is similar to that of the phototube, with electrons being emitted
on exposure to radiation. In place of a single wire anode, however, the PMT has a series of
electrodes called dynodes. The electrons emitted from the cathode are accelerated toward the
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first dynode, which is maintained 90 to 100 V positive with respect to the cathode. Each
accelerated photoelectron that strikes the dynode surface produces several electrons, called
secondary electrons, which are then accelerated to dynode 2, which is held to 90 to 100 V more
positive than dynode l. Again, electron amplification results. By the time this process has been
repeated at each of the dynodes, 10 5 to 10 7 electrons have been produced for each incident
photon. This cascade of electrons is finally collected at the anode to provide an average current
that is further amplified electronically and measured.
Figure: Diagram of a photomultiplier tube: a). photograph b). cross-sectional view; c) electrical
diagram illustrating dynode polarization and photocurrent measurement. Radiation striking the
photosensitive cathode gives rise to photoelectrons by the photoelectric effect. Dynode D1 is
held at a positive voltage with respect to the photocathode. Electrons emitted by the cathode are
attracted to the first dynode and accelerated in the field. Each electron striking dynode D1 thus
gives rise to 2 to 4 secondary electrons. These are attracted to dynode D2, which is again positive
with respect to dynode D1. The resulting amplification at the anode can be 10 6 or greater. The
exact amplification factor depends on the number of dynodes and the voltage difference between
each.
Photoconductive Cells
Photoconductive transducers consist of a thin film of a semiconductor material, such as lead
sulfide, mercury cadmium telluride (MCT), or indium antimonide, deposited often on a
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nonconducting glass surface and sealed in an evacuated envelope. Absorption of radiation by
these materials promotes non-conducting valence electrons to a higher energy state, which
decreases the electrical resistance of the semiconductor. Typically, a photoconductor is placed in
series with a voltage source and a load resistor and the voltage drop across the load resistor
serves as a measure of the radiant power of the beam of radiation. The PbS and InSb detectors
are quite popular in the near-IR region of the spectrum. The MCT detector is useful in the mid-
and far-IR regions when cooled with liquid N 2 to minimize thermal noise.
Silicon Photodiodes and Photodiode Arrays: Crystalline silicon is a semi-conductor, a material
whose electrical conductivity is less than that of a metal but greater than that of an electrical
insulator. Silicon is a Group IV element and thus has four valence electrons. In a silicon crystal,
each of these electrons is combined with electrons from four other silicon atoms to form four
covalent bonds. At room temperature, sufficient thermal agitation occurs in this structure to
liberate an occasional electron from its bonded state, leaving it free to move throughout the
crystal. Thermal excitation of an electron leaves behind a positively charged region termed a
hole, which, like the electron, is also mobile. The mechanism of hole movement is stepwise, with
a bound electron from a neighbouring silicon atom jumping into the electron-deficient region
(the hole) and thereby creating another positive hole in its wake. Conduction in a semiconductor
involves the movement of electrons and holes in opposite directions.
The conductivity of silicon can be greatly enhanced by doping, a process in which a tiny,
controlled amount (approximately 1 ppm) of a Group V or Group III element is distributed
homogeneously throughout a silicon crystal. For example, when a crystal is doped with a Group
V element, such as arsenic, four out of five of the valence electrons of the dopant form covalent
bonds with four silicon atoms, leaving one electron free to conduct. When the silicon is doped
with a Group III element, such as gallium, which has three valence electrons, an excess of holes
develops, which also enhances conductivity. A semiconductor containing unbonded electrons
(negative charges) is termed an n-type semiconductor, and one containing an excess of holes
(positive charges) is a p-type. In an n-type semiconductor, electrons are the majority carrier: in a
p-type, holes are the majority carrier.
Present silicon technology makes it possible to fabricate what is called a pn junction or a pn
diode, which is conductive in one direction and not in the other. Photodiodes are semiconductor
pn-junction devices that respond to incident light by forming electron-hole pairs.
Thermal Detectors
The convenient photon detectors discussed in the previous section cannot be used to measure
infrared radiation because photons of these frequencies lack the energy to cause photoemission
of electrons; as a consequence, thermal detectors must be used. Unfortunately, the performance
characteristics of thermal detectors are much inferior to those of phototubes, photomultiplier
tubes, silicon diodes, and photovoltaic cells. A thermal detector has a tiny blackened surface that
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absorbs infrared radiation and increases in temperature as a consequence. The temperature rise is
converted to an electrical signal that is amplified and measured.
Signal Processors and Readouts
A signal processor is ordinarily an electronic device that amplifies the electrical signal from the
detector; in addition, it may alter the signal from dc to ac (or the reverse), change the phase of
the signal, and filter it to remove unwanted components. The signal processor may also be called
on to perform such mathematical operations on the signal as differentiation, integration, or
conversion to a logarithm. Several types of readout devices are found in modern instruments.
Digital meters, scales of potentiometers, Recorders, Cathode-ray tubes and monitors of
microcomputers are some examples.
Signals, Noise, and the Signal-to-Noise Ratio
The output of an analytical instrument fluctuates in a random way. These fluctuations limit the
precision of the instrument and are the net result of a large number of uncontrolled random
variables in the instrument and in the chemical system under study. An example of such a
variable is the random arrival of photons at the photocathode of a photomultiplier tube. The term
noise is used to describe these fluctuations, and each uncontrolled variable is a noise source. The
average value of the output of an electronic device is called the signal and the standard deviation
of the signal is a measure of the noise. An important figure of merit for analytical instruments,
stereos, compact-disk players, and many other types of electronic devices is the signal-to-noise
ratio (S/N). The signal-to-noise ratio is usually defined as the ratio of the average value of the
output signal to its standard deviation. As modern instruments have become more computerized
and controlled by sophisticated electronic circuits, various methods have been developed to
increase the signal-to-noise ratio of instrument outputs. These methods include analog filtering,
lock-in amplification, boxcar averaging, smoothing, and Fourier transformation.
Optical systems used in spectroscopy: Single beam versus double beam
Instruments of two types are used to measure absorption in optical spectroscopy: Single beam
and double beam spectroscopy.
Single beam spectroscopy: uses one beam of radiation and passes it through a single cell,
generally less expensive. They are primarily used for studies performed at a single wavelength,
i.e. they are not as often used to scan through a wavelength region. The wavelength at which the
study is performed can be varied by adjustment of the monochromator.
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Figure: single- beam Spectroscopy
Double beam spectroscopy: divides the radiation into two beams which are passed through
separate cells. In double beam instruments one of the two cells is filled with either pure solvent
or a blank solution and the second cell is filled with the sample. Since the readout from the
instrument is the difference between the amount of light absorbed in the two cells, the absorption
in the sample is automatically corrected for absorption occurring in the blank/ solvent and for
changes in intensity of the incident radiation with wavelength. Double- beam instruments can be
used at fixed wavelength or they can be used to scan through a wavelength region.
Figure: Double- beam spectroscopy
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