Chapter Three
Instrumentation for optical spectroscopy
➢ Typical spectroscopic instruments that involve absorption techniques contain five
components:
1. a stable source of radiant energy (radiation source)
2. a transparent container for holding the sample (sample holder)
3. a device that isolates a restricted region of the spectrum for measurement
(wavelength selector)
4. a radiation detector, which converts radiant energy to a usable electrical
signal (radiation detector)
5. a signal processor and readout, which displays the transduced signal on
a meter scale, a computer screen, a digital meter, or another recording
device.
➢ These components are almost similar for all absorption techniques with small
variations.
1
To measure the
To emit To isolate a To hold To display
transmitted radiant
stable restricted region the the
energy & convert it
radiation of the spectrum sample transduced
in to a usable
signal
electrical signal
Figure 1: Block diagram of components of instruments for
absorption spectroscopy. It shows the arrangement for absorption
measurements.
In some instruments, the position of the sample & -selector
is reversed
2
Figure 2. The main components of a typical
spectrometer.
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1. Radiation Sources
➢ An ideal radiation source for spectroscopy should have the following characteristics:
1. The source must emit radiation over the entire wavelength range to be studied.
For example, to study in the visible spectrum, the radiation source used should
emit radiation from 350 nm - 800 nm
2. The intensity of radiation over the entire wavelength range must be high
enough so that extensive amplification of the signal from the detector can be
avoided.
[Link] intensity of the source should not vary significantly at different
wavelengths (it should be constant).
[Link] intensity of the source should not fluctuate over long time intervals.
[Link] intensity of the source should not fluctuate over short time intervals. Short
time fluctuation in source intensity is called “flicker”.
➢ This is common and basic component of instrumentation in AAS, NMR, UV–vis and IR
spectroscopy. It produces an emr in the required region.
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The source of radiation can be classified into two class:
Continuum Sources & Line sources
I. Continuum Sources
➢ These are radiation sources that produces radiation over a wide range of
wavelengths, with a relatively smooth variation in intensity as a function of
wavelength.
➢ These are commonly used in UV-vis spectroscopy, IR spectroscopy and NMR.
➢ It produces a radiation in the required region that contains broad wavelengths.
➢ The following table shows common continuum sources that has wide
applications.
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Table 1: Common continuum radiation sources in spectroscopy
Source Wavelength Region Useful for
H2 and D2 lamp (Deuterium continuum source from
UV molecular absorption
lamp) 160–380 nm
continuum source from
Tungsten lamp Vis molecular absorption
320–2400 nm
continuum source from
Xe arc lamp molecular fluorescence
200–1000 nm
continuum source from
Nernst glower IR molecular absorption
0.4–20 mm
continuum source from
Globar IR molecular absorption
1–40 mm
continuum source from
Nichrome wire IR molecular absorption
0.75–20 mm
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II. Line Sources
➢ These are source that emits radiation at only selected wavelengths (very narrow bands of
wavelengths).
➢ It has widely used in atomic absorption and atomic and molecular fluorescence
spectroscopy.
➢ The following table shows commonly used line sources
Table 2: Common Line radiation sources in spectroscopy
Source Wavelength Region Useful for
hollow cathode lamp line source in UV/Vis atomic absorption (AAS)
Hg vapor lamp line source in UV/Vis molecular fluorescence
atomic and molecular
laser line source in UV/Vis absorption, fluorescence
and scattering
7
Figure 3. Emission spectrum from
a typical continuum source
Figure 4. Emission spectrum
from a typical line source
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2. Wavelength Selectors
➢are devices used to filter out the unwanted wavelengths and allow only the wavelength
of interest to pass. i.e., for selecting the wavelength of maximum absorption
➢Ideally, the out put from a wave length selector would be radiation of a single
wavelength or frequency. However, no real wavelength selector even approaches this
ideal. Instead a band as shown in figure 5.
➢Nominal wavelength: is the principal wavelength provided by a wavelength selection
device.
We can not isolate a single
wavelength of radiation from a
continuum source. Instead, a
selector passes a narrow band of
radiation characterized by a nominal
(effective band width)
Figure 5. Band of radiation
exiting wavelength selector
showing the nominal wave
length & effective band pass
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➢ The electromagnetic spectrum is a continuous wavelength band. Thus
wavelength selection in a spectrochemical instrument actually consists of the
selection of a narrow wavelength band from the larger band.
For example, isolating a band of 400 nm-450 nm from a band of 350nm-800nm.
➢ The width of the band that is allowed to pass is called the bandwidth.
➢ The narrowness of the band that is allowed to pass varies from one design to
another and is called the resolution.
➢ Thus a high resolution (narrow bandwidth) is ideal, although many applications
do not require the best available resolution and money is often saved by
purchasing instruments with a low resolution (wide bandwidth).
➢ There are two general types of wavelength selectors:
Filters & Monochromators
\\\\\\
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I. Filters
➢ Are wavelength selectors & there are two types of filters
Absorption filters & Interference filters
➢ This means that absorption or interference mechanism is used to control or isolate the
range of selected wavelengths.
a. Absorption filters
➢ The most common type consists of colored glass or of a dye suspended in gelatin and
sandwiched between glass plates.
➢ Absorption filters can be as simple as a piece of colored glass.
For example, a blue glass transmits blue wavelengths of the visible spectrum but absorb
yellow wavelengths. So blue glass can be used to isolate blue region of a visible
radiation.
➢ Colored glass absorption filters can be purchased that isolates various ranges of visible
light.
➢ These filters are stable, simple, and cheap, so they are excellent for use in portable
spectrometers designed to be carried into the field.
➢ limitations
✓ the range of wavelengths transmitted is broad compared with prisms and gratings
(the transmission range may be 50–300 nm for typical absorption filters)
✓ Absorption filters are limited to the visible region of the spectrum and the X-ray
region.
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➢ The color of a substance depends on which wavelength of light it
absorbs.
For example, a solution containing copper ions is blue because it absorbs
the complementary color yellow from white light and transmits the
remaining blue light.
➢ The more concentrated the copper solution, the more yellow light is
absorbed and the deeper the resulting blue color of the solution
➢ In a spectrometric method, the amount of light absorbed would be
measured and related to the concentration
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Table 3. complementary colors
Color absorbed Color observed Absorbed radiation ,
(transmitted color) (nm)
Violet Yellow- green 400-435
Blue Yellow 435-480
Green Purple 500-560
Yellow Blue 580-595
Orange Green-blue 595-605
Red Blue-green 605-750
Green-blue Orange 480-490
Blue-green Red 490-500
Yellow-green violet 560-580
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b. Interference Filters
➢ the interference filter is constructed of multiple layers of different materials.
➢ The filter operates on the principle of constructive interference to transmit selected
wavelength ranges.
➢ The wavelengths transmitted are controlled by the thickness and refractive index of
the center layer of material.
➢ Interference filters can be constructed for transmission of light in the IR, visible, and
UV regions of the spectrum.
➢ Advantages:
✓ The wavelength ranges transmitted are much smaller than for
absorption filters, generally 1–10 nm, and
✓ the amount of light transmitted is generally higher than for absorption
filters.
➢ Limitations of an absorption or interference filter:
✓ 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.
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II. Monochromators
➢ For many spectroscopic methods, it is necessary or desirable to be able to continuously
vary the wavelength of radiation over a broad range. This process is called scanning a
spectrum.
➢ An alternative approach to wavelength selection, which provides for a continuous
variation of wavelength, is the monochromator.
➢ The word “monochromator” is derived from the Latin language, “mono” meaning “one”
and “chromo” meaning “color.”
➢ Monochromators :
✓ are more sophisticated than an absorption filter
✓ Isolates the narrow band of wavelengths from visible and ultraviolet sources.
✓ Are designed for spectral scanning
➢ Spectral scanning implies changing the wavelength of radiation continuously over a
considerable range
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Components of a monochromator
[Link] entrance slit
✓ is a small circular or rectangular hole cut in an otherwise opaque plate, such as a
black metal plate.
✓ The size of the opening is often variable (a variable slit width )
✓ The entrance slit is where light enters the monochromator from the source.
✓ Its purpose is to create a unidirectional beam of light of appropriate intensity
from the multidirectional light emanating from the source.
✓ Its slit width is usually variable so that the intensity of the beam can be adjusted,
the wider the opening the more intense the beam.
✓ The entrance slit allows light from the source to fall on the dispersion element
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2. a dispersing element
➢ The dispersing element disperses the light into its component wavelengths.
➢ The function of the dispersion element is to spread out in space, or disperse, the
radiation falling on it according to wavelength.
➢ There are two types of dispersing element: Prisms & Gratings
i) Prisms
➢ A prism is a three-dimensional triangularly shaped glass or quartz block.
➢ When the light beam strikes one of the three faces of the prism, the light emerging through
another face is dispersed.
➢ When electromagnetic radiation passes through a prism, it is refracted because of the difference
in the refractive index of the material in which the prism is made up of and the air.
➢ The refractive index depends on the wavelength and, therefore, so does the degree of refraction.
➢ Shorter wavelengths are refracted more than longer wavelengths.
➢ The effect of refraction is to “spread” the wavelengths apart into different wavelengths.
➢ By rotation of the prism, different wavelengths of the spectrum can be made to pass through an
exit slit and the sample.
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➢For example, When a white light passes through a prism it will disperse
into its components (the seven rainbow colors) containing violet, indigo,
blue, green, yellow, orange and red (VIBGYOR)
➢The dispersion element is rotated slowly, allowing first violet light through
the exit slit, then blue light, and so on all the way to red light.
➢In this way, the monochromator sorts polychromatic radiation from a
source into nearly monochromatic radiation leaving the exit slit.
18 Figure 5: Dispersion of visible light by a prism.
➢ Prisms are used to disperse IR, visible, and UV radiation.
➢ However, because of its non linear dispersion, it works more effectively for the shorter
wavelengths
➢ Prisms and lenses that are made up of glasses can be used to disperse visible radiation.
➢ Quartz or fused silica must be used in the ultraviolet region.
➢ To disperse IR we use prisms made up of crystals of alkali halides since they are
transparent in the IR region. Sodium chloride (rock salt) is used in most instruments and
is useful for the entire region from 2.5 to 15.4 µm (4000 to 650 cm-1).
➢ For longer wavelengths, KBr (20 to 25 µm) or CsI (10 to 38 µm) can be used. Due to
solubility effect they must be kept dry.
ii) Gratings
➢ A diffraction grating (reflection) is made by ruling on a polished metal surface (such as
Al)
➢ A grating is a reflective optical component with a series of closely spaced, parallel ruled
19 grooves
➢ Athin protective layer of silica (SiO2) on top of the Al protects the metal surface from
oxidising (since oxidising would reduce its reflectivity)
➢ When light is reflected from this surface,that which strikes the rulings is disspated by
scattering;the unruled portions reflect regularly,acting as individual light sources.
➢ i.e.,rulings (lines) used for scattering while the unruled surface is used for reflecting
➢ Overlaping of the waves from these sources establishes an interference pattern which
results in the dispersion of the reflected light in to its component wave lengths.
➢ A diffraction grating consists of a series of closely spaced parallel grooves cut (or ruled)
into a hard glass, metallic, or ceramic surface .
➢ The surface may be flat or concave, and is usually coated on the ruled surface with a
reflective coating.
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➢UV, visible, and IR radiation can be dispersed by a diffraction grating.
➢A grating for use in the UV and visible regions will contain between 500 and 5000
grooves/mm, while a grating for the IR region will have between 50 and 200 grooves/mm.
Collimating lens (in the
case of prism) & concave
mirror (in the case of
grating) are used to produce
parallel beam of radiation
collimation
Figure 6: Types of
monochromators:
(a) grating
monochromator:
(b) (b) prism
monochromator
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3. An exit slit
➢ Is used to isolate the desired spectral band by blocking all of the dispersed radiation
except that in the desired range
➢ The exit slit width can be variable too, but making it wider would result in a wider
wavelength band (a wider bandwidth) passing through, which can be undesirable.
➢ Generally,polychromatic radiation enters the monochromator through the entrance
[Link] beam is collimated, and then strike the dispersing element at an [Link] beam
is split (dispersed) in to its component wavelength by the grating or prism. By
mechanically moving (or rotating) the dispersing element (grating or prism) or the exit
slit. Radiation of only a particular wavelength leaves the monochromator through the
exit slit & is passed on to the sample compartment to pass through the sample.
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3. Sample Holder
➢ In molecular spectroscopy we usually use sample holder.
➢ They are used to hold the sample.
➢ They are placed after the wavelength selector and before the detector.
➢ In some instruments it is placed between the radiation source and the
wavelength selector.
➢ They usually are called cells or cuvettes.
➢ The sample holder can be made up of different substances depending on the type
of the radiation being used.
➢ The major criterion for a material to be used as a sample holder is that it must be
transparent in the region of radiation used.
In the visible region:
✓ Silica glass is ordinarily used (in the region from 375 to 2000 nm region
because of its low cost compared with quartz)
✓ Plastic containers can also be used
✓ Quartz can be used, but it is expensive
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➢ Sample holders made up of quartz can be used in UV-region.
➢ Glasses and quartz are not fairly transparent in the IR region. So they are not usually used
in the IR spectroscopy.
➢ For IR region sample containers that are made up of crystal of alkali halides are usually
used.
For example, NaCl, KBr, CaF2
➢ 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.
➢ The quality of spectroscopic data is critically dependent on the way that cells are used
and maintained.
➢ Finger prints, 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.
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▪ 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
Figure 7: Typical examples
of commercially available
cells for the UV/visible
region.
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4. Detector (radiation transducer)
➢ is a device that converts radiation energy into electrical signal.
➢ The simplest and first detector to obtain spectroscopic information is the human eye; 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.
➢ Invariably in modern instruments, the information of interest is encoded and processed
as an electrical signal.
properties of the Ideal Transducer
The ideal transducer
✓ would have a high sensitivity
✓ would have a high signal-to-noise ratio
✓ would have a constant response over a considerable range of wavelengths.
✓ would exhibit a fast response time and a zero output signal in the absence of
illumination,
➢ The electrical signal produced by the ideal transducer would be directly proportional to
the radiant power P. That is, S = kP, where S is the current or voltage output of the
transducer, and k is the calibration sensitivity
S = kP + kd, where kd represents the dark current, which is usually constant over short
measurement periods. Instruments with transducers that produce a dark current are Often
26 equipped with a compensating circuit that reduces kd to zero.
Types of Radiation Transducers
❑ There are two general types of radiation transducers;
a) Photon transducers & b)Thermal transducers
a) Photon transducers
We have two general types under this category.
i) Photocurrent transducers
ii) Photoconductive transducers
i) Photocurrent transducers
➢ these transducers have an active surface usually metals that absorbs radiation.
➢ The absorption of the radiation energy causes emission of electrons resulting a current
usually called photocurrent.
➢ The current produced is directly proportional to the power of radiation that strikes the
active surface.
➢ The generated current is measured and converted into a form that is perceived by us.
➢ Photovoltaic cells, phototubes, photomultiplier tubes, silicon photodiodes are an example
of photocurrent transducers.
➢ Photocurrent transducers are widely used in the UV visible region. They are not used in
the IR region. IR has low energy and it cannot generate a current that results no detection.
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ii) Photoconductive transducers
➢ In this types of transducers the absorption of a radiation by the transducer
decreases the resistance or increases its conductivity.
➢ The change in the resistance is measured. The change in the resistance is
proportional to the power of radiation.
➢ The most sensitive transducers for monitoring radiation in the near-
infrared region (0.75 to 3 µm) are semiconductors whose resistances
decrease when they absorb radiation within this range.
➢ Absorption of radiation by these materials promotes some of their bound
electrons into an energy state in which they are free to conduct
electricity. The resulting change in conductivity can then be measured.
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b) Thermal Transducers
➢ In thermal transducers, the radiation impinges on and is absorbed by a small blackbody
and results in an increase in temperature; the resultant temperature rise is measured.
➢ The problem of measuring infrared radiation by thermal means is compounded by
thermal noise from the surroundings.
➢ For this reason, thermal detectors are housed in a vacuum and are carefully shielded
from thermal radiation emitted by other nearby objects.
➢ To further minimize the effects of extraneous heat sources, the beam from the source is
generally chopped.
➢ In this way, the analyte signal, after transduction, has the frequency of the chopper and
can be separated electronically from extraneous noise, which usually varies slowly
withtime.
➢ Thermocouples, bolometers and pyroelectric transducers are examples of thermal
detectors.
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Table 4. Examples of Photon transducers & Thermal transducers
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5. Signal Processors and Readouts
✓ The signal processor is usually an electronic device that amplifies the
electrical signal from the transducer.
✓ In addition, it may alter the signal from de to ac (or the reverse), change
the phase of the signal, and filter it to remove unwanted components.
✓ Furthermore, the signal processor may 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.
✓ Some of these devices include digital meters, recorders, cathode ray
tubes, and computer displays.
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Figure 8: Basic structure of spectrophotometers
32
Optical systems used in spectroscopy:
single beam versus double beam instruments
▪ A spectrophotometer is an instrument that analyzes the components of
a solution by observing the capability to absorb light. There are two
main types of spectrophotometers; single beam and double beam
spectrophotometer.
▪ Single beam spectrophotometer is an analytical instrument in which all
the light waves coming from the light source passes through the
sample.
▪ Therefore, the measurements are taken as the intensity of light before
and after the light pass through the sample.
▪ A single beam spectrophotometer measures the concentration of an
analyte in a sample by measuring the amount of light absorbed by that
analyte. Here, the Beer Lambert Law comes into operation. This law
states that the concentration of an analyte is directly proportional to the
absorbance.
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Figure 9: Single Beam Spectrophotometer
34
Advantages of a Single Beam Spectrophotometer
✓ These single beam spectrophotometers are more compact and
optically simpler than double beam spectrophotometers.
✓ Cost-Effective: Single beam instruments are less expensive as
compared to the other alternative.
✓ Better Performance: High energy throughput due to the non-
splitting of the source beam results in high sensitivity of
detection.
❑ The sensitivity of detection of the light beam after it passes
through the sample is high since it uses a non-split light beam
(therefore, high energy exists throughout). Single beam
spectrophotometers are available in analysis at visible and
ultraviolet wavelength ranges.
Disadvantages of a Single Beam Spectrophotometer
✓ Instability: This happens due to lack of compensation for disturbances
like electronic circuit fluctuations, voltage fluctuations, mechanical
component’s instability, or drift in the energy of light sources. Such
35
drifts cause abnormal fluctuations in the results.
Double Beam Spectrophotometer
▪ Double beam spectrophotometer is an analytical instrument
in which the light beam coming from the light source splits
into two fractions. One fraction acts as the reference (the
reference beam) while the other fraction passes through the
sample (sample beam). As a result, the reference beam does
not pass through the sample.
▪ Unlike the single beam variant, this one does not standardize
or blank the instrument before use. Instead, it splits the beam
into two parts for the same purpose. One of the beam parts is
passed through the given object while the other passes
through a reference standard.
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Figure 10: The Pathway of Light Beam in a Double Beam
Spectrophotometer
37
Working:
▪ The light beam from the source is split into sample beam and reference
beam by the mechanical chopper. The reference beam monitors the lamp
energy whereas the sample beam reflects sample absorption.
▪ The observed absorbance measurement is the ratio of the sample and
reference beams which are recombined before moving to the
monochromator. This arrangement compensates the effects due to drift in
lamp intensity, electronic and mechanical fluctuations which affect both
the sample and reference beams equally.
Advantages of a Double Beam Spectrophotometer
✓ More Reliable Detection: Modern improvements in optics
permit a high level of automation and offer the same or even better level
of detection as compared to earlier single beam systems. Instability
factors due to lamp drift, stray light, voltage fluctuations do not affect the
measurement in real-time.
✓ No Warm-Up Time: Little or no lamp warm-up time is required. This
not only improves the throughput of results but also conserves lamp life.
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Disadvantages of a Double Beam Spectrophotometer
▪ Cost:The number of components get increased in this
[Link] a more significant price will be
associated with the more complex structured instrument.
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Single Beam vs Double Beam Spectrophotometer
Single beam spectrophotometer is an Double beam spectrophotometer is an
analytical instrument in which all the analytical instrument in which the light
light waves coming from the light beam coming from the light source
source passes through the sample. splits into two fractions.
Light Beam
Double beam spectrophotometer uses a
Single beam spectrophotometer uses a light beam that is split into two
non-split light beam. fractions before passing through the
sample.
Measurement
The measurements taken from double
The measurements taken from single
beam spectrophotometers are highly
beam spectrophotometers are less
reproducible because electronic and
reproducible because a single light
mechanical effects on both sample and
beam is used.
reference beams are equal.
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