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
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
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, and line
sources.
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
2000–3400K. 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 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 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
Light Sources for Atomic Absorption
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.
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
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: less expensive than interference filters and 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 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 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:
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.
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-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.
[Link] 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.
[Link] prisms: designed to change direction of propagation of
beam, orientation, or both
[Link] 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
region (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 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.
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 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: include phototubes, photomultiplier tubes, silicon
photodiodes, and photodiode arrays.
Phototubes and Photomultiplier Tubes: The response 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.
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 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.
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 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
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. 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. 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. The
term noise is used to describe these fluctuations, and each uncontrolled variable
is a noise source. 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.
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.