Chapter one

Spectroscopy and Electromagnetic Radiation

Introduction
Measurements based on light and other forms of electromagnetic radiation are widely used
throughout analytical chemistry. Spectroscopy is the science which deals with the interactions of
radiation and matter. Spectroscopic analytical methods are based on measuring the amount of
radiation produced or absorbed by molecular or atomic species of interest. Spectroscopic
methods can be classified according to the region of the electromagnetic spectrum involved in
the measurement. The regions of the spectrum that have been used include γ-ray, X-ray,
ultraviolet (UV), visible, infrared (IR), microwave, and radiofrequency (RF). Spectroscopy has
played a vital role in the development of modem atomic theory. In addition, spectrochemical
methods have provided perhaps the most widely used tools for the elucidation of molecular
structure as well as the quantitative and qualitative determination of both inorganic and organic
compounds.
Electromagnetic radiation and its property
Electromagnetic radiation is a form of radiant energy that is propagated as a transverse wave.
Electromagnetic radiation is a form of energy that is transmitted through space at enormous
velocities. Electromagnetic radiation is the means for many of our interactions with the world:
light allows us to see; radio waves give us TV and radio; microwaves are used in radar
communications; X-rays allow glimpses of our internal organs. Electromagnetic radiation is the
messenger, or the signal from sender to receiver. The sender could be a TV station, a star, or the
burner on a stove. The receiver could be a TV set, an eye, or an X-ray film. In each case, the
sender gives off or reflects some kind of electromagnetic radiation. Electromagnetic radiation, or
light, is a form of energy whose behavior is described by the properties of both waves and
particles. The optical properties of electromagnetic radiation, such as diffraction, are explained
best by describing light as a wave. Many of the interactions between electromagnetic radiation
and matter, such as absorption and emission, however, are better described by treating light as a
particle, or photon (discrete packets of energy). In contrast to sound waves, light requires no
supporting medium for its transmission; thus, it easily passes through a vacuum. Light also
travels nearly a million times faster than sound.
Wave Properties
In dealing with phenomena such as reflection, refraction, interference, and diffraction,
electromagnetic radiation is conveniently modeled as waves consisting of perpendicularly
oscillating electric and magnetic fields. The electric field for a single-frequency wave oscillates
sinusoidally in space and time. Electromagnetic radiation consists of oscillating electric and
magnetic fields that propagate through space along a linear path and with a constant velocity.
The electric field is represented as a vector whose length is proportional to the field strength.
Oscillations in the electric and magnetic fields are perpendicular to each other, and to the
direction of the wave’s propagation.

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Fig.1.Wave nature of Electromagnetic radiation; Plane-polarized electromagnetic radiation
showing the electric field, the magnetic field, and the direction of propagation.
Wave Characteristics

The wave is described either in terms of its wavelength (the distance of one complete cycle) or in
terms of the frequency (the number of cycles passing a fixed point per unit time). The reciprocal
of the wavelength is called the wavenumber (the number of waves in a unit length or distance per
cycle). The relationship between the wavelength and frequency is

 wavelength in centimetre

v- frequency in reciprocal seconds

c – velocity of light = 3x1010cm/s

Wavenumber, the reciprocal of wavelength is,

The amplitude(A) of an electromagnetic wave is a vector quantity that provides a measure of

the electric or magnetic field strength at a maximum point in the wave.

The period of an electromagnetic wave is the time in seconds that it takes for successive
maxima or minima to pass a point in space. The frequency, v, is the number of oscillations of the
electric field vector per unit time and is equal to l/p. The frequency of an electromagnetic wave is
the number of oscillations that occur in 1 second. The unit of frequency is the hertz (Hz) which
corresponds to one cycle per second. The frequency of a beam of electromagnetic radiation does
not change as it passes through different media. The frequency of a light wave, or any wave of
electromagnetic radiation, is determined by the source that emits it and remains constant
regardless of the medium traversed. In contrast, the velocity, v, of the wave front through a
medium depends on both the medium and the frequency.
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Fig.2. Wave properties of Electromagnetic radiation

The Speed of Light

In a vacuum light travels at its maximum velocity. This velocity, which is given the special
symbol c, is 2.99792 x 10 8 ms-1. The velocity of light in air is only about 0.03% less than its
velocity in a vacuum. Thus, for a vacuum or for air,

c = vλ = 3.00 x 10 8 m s-1 = 3.00 x 1010 cm s-1

In a medium containing matter, light travels with a velocity less than c because of interaction
between the electromagnetic field and electrons in the atoms or molecules of the medium. Since
the frequency of the radiation is constant, the wavelength must decrease as the light passes from
a vacuum to a medium containing matter. This effect is illustrated in the Figure below for a beam
of visible radiation.

Fig.3. Change in wavelength as radiation passes from air into a dense glass and back to air. Note
that the wavelength shortens by nearly 200 nm or more than 30% as it passes into glass: a reverse
change occurs as the radiation again enters air.

The refractive index (η)of a medium measures the extent of interaction between
electromagnetic radiation and the medium through which it passes. It is defined by η = c/v. For
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example, the refractive index of water at room temperature is 1.33, which means that radiation
passes through water at a rate of c/1.33 or 2.26 x 1010 cm s -l. In other words, light travels 1.33
times slower in water than it does in a vacuum. The velocity and wavelength of radiation become
proportionally smaller as the radiation passes from a vacuum or from air to a denser medium,
while the frequency remains constant.

The Particle Nature of light: Photons

In many radiation/matter interactions, it is most useful to consider light as consisting of photons

or quanta. Electromagnetic radiation possesses a certain amount of energy. When a matter
absorbs electromagnetic radiation it undergoes a change in energy. When a photon is absorbed
by a sample, it is “destroyed,” and its energy acquired by the sample. The energy of a unit of
radiation, called the photon, is related to the frequency or wavelength by

E- energy of the photons in ergs

h- planck’s constant= 6.62x10-34J.S

The shorter the wavelength or the greater the frequency, the greater the energy.

Example: Calculate the frequency and wavenumber of a beam of infrared radiation with a
wavelength of 5.00 µm. Calculate the energy in joules of One photon of the radiation.

Radiant Power and Intensity

The radiant power P in watts (W) is the energy of a beam that reaches a given area per unit time.
The intensity is the radiant power-per-unit solid angle. Both quantities are proportional to the
square of the amplitude of the electric field. Although it is not strictly correct, "radiant power"
and "intensity" are frequently used interchangeably.

Interaction of radiation and matter

The most interesting types of interactions in spectroscopy involve transitions between different
energy levels of chemical species. As a molecule interact with electromagnetic radiation, the
internal energy of the molecule increase, the increase in energy being equal to the energy of the
absorbed radiation (hv) and there will be transition from lower energy state to higher energy
state. Electromagnetic radiation only interacts with molecules at specific energies. In order for
the electric field to induce a dipole, the energy difference between the two states must be equal to
the energy contained in the photon, and the orientation of the electric field vector must match the
orientation of the molecule. (The mathematical derivation of this is very complex; instead, we
will consider the interaction with light using a more qualitative approach.)
The state of a molecule can be described by an energy diagram such as the one shown below.
The energy diagram shows two electronic states, with each electronic state comprised of a
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variety of vibrational and rotational states. The difference in energy between one rotational state
and the next is 4 kJ/mol, which is in the range of thermal energy of the molecule, allowing
molecules to adopt more than one rotational mode. However, the vibration energy levels are
about 40 kJ/mol apart, and electronic states are 150 to 450 kJ/mol apart (depending on the
molecule).

Each of the energy states the molecule can adopt is described by a wavefunction. The lowest
energy state, the ground state, is ψ. Other states within the ground electronic state are ψ 0,v,r where
the v and r subscripts refer to the specific vibrational and rotational states. The interaction of a
molecule with electromagnetic radiation in the ultraviolet or visible energy range induces a
transition from one electronic state to another.

Rotational transition - the molecule rotates about various axes, the energy of rotation being at
definite energy levels, so the molecule may absorb radiation and be raised to a higher rotational
energy level.

Vibrational transition – the atoms or groups of atoms within a molecule vibrate relative to each
other and the energy of this vibration occurs at definite quantized levels. The molecule may then
absorb a discrete amount of energy and be raised to a higher vibrational energy level.

Electronic Transition –If EMR of an energy identical with the difference between the ground
state of the molecule and an excited electron state strikes the molecule, the electron can absorb
the energy from the EMR and jump from the ground level to excited level and this is called
electronic transition.

The relative energy levels of the three transition processes are in the order :

Electronic > vibrational > Rotational

Absorption and Emission of Radiation

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The interaction of the light (electro-magnetic radiation) with a substance and the subsequent
energy transfer ends with three main processes namely:

Absorption: The process by which the energy of the light (in the form of photons) is
transferred to the atom or molecule raising them from the ground state to an excited state
Fluorescence: The absorbed energy is rapidly lost to the surroundings by collisions within the
system and relax back to the ground state. Sometimes the energy is not lost in this way but is re-
emitted a few milli seconds later, which is referred as fluorescence. In fluorescence materials
emit light of different energies (or wavelengths) than they absorb.
Some materials absorb, save the energy for long times, and so give off photons long after the
energy source has gone. This delayed emission of light is phosphorescence; you’ve all seen
phosphorescence as “glow in the dark” stickers, T-shirts, Frisbees, etc.
Emission: If the substances (atoms or molecules) are heated to high temperatures (in a flame or
in an electric discharge) the electrons are exited to higher energy levels. Later, they relax to the
ground state with the emission of radiation, the magnitude of which is more or less equivalent to
absorbed energy.
Most of the analytical techniques are based on the light interactions with the substances and
utilise any of the above associated processes.
Electromagnetic Spectrum
The electromagnetic spectrum is arbitrarly broken down into different regions according to
wavelength. The electromagnetic spectrum covers an enormous range of energies (frequencies)
and thus wavelengths. Useful frequencies vary from > 10 19 Hz (γ-ray) to 103Hz (radio waves).
An X-ray photon (v = 3x10 18 Hz, λ = 10-10 m), for example, is approximately 10,000 times as
energetic as a photon emitted by an ordinary light bulb (v = 3x10 14Hz, λ = 10-6m) and 1015 times
as energetic as a radio-frequency photon (v = 3x103Hz, λ = 10 -5 m).

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Fig.5. Electromagnetic spectrum

Fig.4. Interaction of an analyte with electromagnetic radiation can result in the types of changes.
Note that changes of electron distribution occur in the UV/visible region.

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Spectroscopic Measurements
Spectroscopists use the interactions of radiation with matter to obtain information about a
sample. The sample is usually stimulated in some way by applying energy in the form of heat,
electrical energy, light, particles, or a chemical reaction. Prior to applying the stimulus, the
analyte is predominantly in its lowest energy state or ground state. The stimulus then causes
some of the analyte species to undergo a transition to a higher energy or excited state. We
acquire information about the analyte by measuring the electromagnetic radiation emitted as it
returns to the ground state or by measuring the amount of electromagnetic radiation absorbed. In
emission spectroscopy, the intensity of emitted radiation measured and in Absorption
spectroscopy the amount of absorbed radiation measured and these related to the properties of
analyte.

Figure: Measurement of Emitted Radiation

Figure: Measurement of Absorbed radiation

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Every molecular species is capable of absorbing its own characteristic frequencies of
electromagnetic radiation. This process transfers energy to the molecule and results in a decrease
in the intensity of the incident electromagnetic radiation. Absorption of the radiation thus
attenuates the beam in accordance with the absorption law. In spectroscopy, to attenuate means
to decrease the energy per unit area of a beam of radiation. In terms of the photon model, to
attenuate means to decrease the number of photons per second in the beam.

The absorption law, also known as the Beer-Lambert law or just Beer's law, tells us
quantitatively how the amount of attenuation depends on the concentration of the absorbing
molecules and the path length over which absorption occurs. As light traverses a medium
containing an absorbing analyte, decreases in intensity occur as the analyte becomes excited. For
an analyte solution of a given concentration, the longer the length of the medium through which
the light passes (path length of light), the more absorbers are in the path, and the greater the
attenuation. Also, for a given path length of the light, the higher the concentration of absorbers,
the stronger the attenuation. The following Figure depicts the attenuation of a parallel beam of
monochromatic radiation as it passes through an absorbing solution of thickness b centimeters
and concentration c moles per liter. Because of interactions between the photons and absorbing
particles, the radiant power of the beam decreases from Po to P. The transmittance T of the
solution is the fraction of incident radiation to transmitted by the solution. Transmittance is often
expressed as a percentage called the percent transmittance.

(Transmittance) T = and Percent transmittance, %T = x100%

Absorbance, A is defined as the negative logarithm of the transmittance, and you will note that
absorbance and transmittance bear an inverse relationship.
A = -logT = log

If you have two colored solutions, you may deduce that the darker colored green solution appears
darker because it absorbs more of the light falling on it. Because the darker solution is also the
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more concentrated one, you can also say that the more concentrated one absorbs more of the
light. That is, the absorbance increases as concentration increases.
Next, suppose that there are two test tubes, both containing the same solution at the same
concentration. The only difference is that one of the test tubes is thicker than the other.

We shine light of the same intensity (Po) on both containers. In the first case the light has to
travel through only a short distance, whereas in the second case it has to pass through a much
longer length of the sample. We might deduce that in the second case more of the light will be
absorbed or cut off, since the path length is longer. In other words, absorbance increases as
pathlength increases.
The two observations described above (those dealing with the relationship between absorbance
and concentration and absorbance and path length) constitute the Beer-Lambert law.
Beer-Lambert Law:
Absorbance(A) ∝ path length (b)x concentration(C)
A = abC
where
A(absorbance) is a dimensionless number.
Here, a is a proportionality constant called the absorptivity. Because absorbance is a unitless
quantity, the absorptivity must have units that cancel the units of b and C. If, for example, C has
the units of g L -1 and b has the units of cm, absorptivity has the units of L g -1 cm-1. When we
express the concentration in moles per liter and b in centimeters, the proportionality constant is
called the molar absorptivity and is given the special symbol, ɛ. Thus,
A = ɛbc
ɛ the proportionality constant, is called the molar extinction coefficient or molar absorptivity. It
is a constant for a given substance, provided the temperature and wavelength are constant. It has
units of liter/mol.cm.

Beer’s Law and Multicomponent Samples

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Beer’s Law can be extended to samples containing several absorbing components provided that
there are no interactions between the components. Individual absorbances, A, are additive. For a
two component mixture of X and Y, the total absorbance, Atot, is

Atot = Ax + Ay = εxbCx + εybCy

Generalizing the absorbance for a mixture of n components, Am is given as

Am = =

Spectrophotometric Analysis
The quantitative measurement of light absorption as a function of wavelength can establish both
the identity and the concentration of a substance in solution. The spectrophotometer is an
instrument that separates electromagnetic radiation into its component wavelengths and
selectively measures the intensity of radiation after passing through a sample. A general
approach for spectrophotometric analysis is first finding the absorption spectrum of “finger
prints” of a substance and then determining its concentration.

1. Plotting Absorption Spectra

Recall that the extinction coefficient for any given substance is a constant only so long as the
wavelength of light is constant. You will see that the absorbance changes with wavelength. The
plot of a sample's absorbance of light at various wavelengths is called its absorption spectrum.
(The abscissa or horizontal axis may be expressed in terms of wavelength and the ordinate or
vertical axis in terms of absorbancy.) The plot below gives the absorption spectrum of potassium
permanganate (KMnO4), a purple colored solution, at two different concentrations. Curves 1 and
2 represent the absorption spectra measured under the same conditions except that curve 1
represents a more concentrated solution than curve 2. Note the similar shapes of the curves.

Fig:The absorption spectrum of solutions of potassium permanganate (KMnO4) at two different

concentrations. The solution for curve 1 has a higher concentration than that for curve 2.

2. Choice of Wavelength
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According to the Beer-Lambert Law absorbance is proportional to concentration at each
wavelength. Theoretically we could choose any wavelength for quantitative estimations of
concentration. However, the magnitude of the absorbancy is important, especially when you are
trying to detect very small amounts of material. In the spectra above note that the distance
between curves 1 and 2 is at a maximum at 525 nm, and at this wavelength the change in
absorbance is greatest for a given change in concentration. That is, the measurement of
concentration as a function of concentration is most sensitive at this wavelength. For this reason
we generally select the wavelength of maximum absorbance for a given sample and use it in our
absorbance measurements. Suppose instead that we had chosen a wavelength of 500 nm for our
measurement, this wavelength being on one of the steep portions of the curve. Examination of
the curve shows that even a small fluctuation in the wavelength will cause a large error in the
absorbance. Most spectrophotometers show a slight fluctuation in the wavelength, so errors in
absorbance will be magnified if we select a wavelength such as 500 nm in our preceding
example.
3. Plotting Calibration Graphs
Once we have chosen the correct wavelength, the next step is to construct a calibration curve or
calibration plot. This consists of a plot of absorbance versus concentration for a series of
standard solutions whose concentrations are accurately known. Because calibration curves are
used in reading of the unknown concentrations, their accuracy is of absolute importance.
Therefore, make the standard solutions as accurately as possible and measure their absorbances
carefully. Each standard solution should be prepared in identically the same fashion, the only
difference between them being their concentrations. Slope of the best straight line through the
data points in the calibration plot is
Slope = Δ (absorbance)
Δ (concentration)
The Beer-Lambert Law (A = abC) implies that when concentration is equal to zero (C = 0),
absorbance must also be zero (A = 0). In other words, the calibration line must pass through the
origin.

Limitations to Beer’s Law

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According to Beer’s law, a calibration curve of absorbance versus the concentration of analyte in
a series of standard solutions should be a straight line with an intercept of 0 and a slope of ab or
εb. In some cases, deviation from linearity can occur and this is divided into three categories:
Fundamental, Chemical and Instrumental deviation.
1. Fundamental Limitation to Beer’s law
Beer’s law is a limitating law that is valid only for low concentration of analyte. There are two
contributions to this fundamental limitations to Beer’s Law. The first is that at higher
concentrations the individual particles of analyte no longer behave independently of one another.
The resulting interaction between particles of analyte may change the value of ε.
A second contribution is that the absorptivity a and molar absorotivity ε, depends on the
sample’s refractive index. Since the refractive index varies with the analyte’s concentration, the
value of a and ε will change.
2. Chemical Limitations to Beer’s Law
Chemical deviations from Beer’s Law can occur when the absorbing species is involved in an
equilibrium reaction. Consider as an example an analysis for the weak acid, HA. To construct a
Beer’s law calibration curve, several standards containing known total concentration of HA, C tot,
are prepared and the absorbance of each is measured at the same wavelength. Since HA is a
weak acid, it exists in equilibrium with its conjugate weak base, A-

If both HA and A– absorb at the selected wavelength, then Beers law is written as
A = εHAbCHA + εAbCA
where CHA and CA are the equilibrium concentrations of HA and A–. Since the weak
acid’s total concentration, Ctot, is
Ctot = CHA + CA
–
the concentrations of HA and A can be written as
CHA = aHACtot

CA = (1 – aHA)Ctot

where aHA is the fraction of weak acid present as HA. Substituting and rearranging gives

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Because values of εHA may depend on the concentration of HA, the above equation may not be
linear. A Beer’s law calibration curve of A versus Ctot will be linear if one of two conditions is
met. If the wavelength is chosen such that εHA and εA are equal, then the equation simplifies to

A = εAbCtot

and a linear Beer’s law calibration curve is realized. Alternatively, if ε HA is held constant for all
standards, it will be a straight line at all wavelengths. Because HA is a weak acid, values of ε HA
change with pH. To maintain a constant value for ε HA, therefore, we need to buffer each standard
solution to the same pH. Depending on the relative values of ε HA and εA the calibration curve will
show a positive or negative deviation from Beer’s law if the standards are not buffered to the
same pH.

3. Instrumental Limitations to Beer’s Law

There are two principal instrumental limitations to Beer’s law. The first limitation is that Beer’s
law is strictly valid for purely monochromatic radiation; that is, for radiation consisting of only
one wavelength. However, even the best wavelength selector passes radiation with a small, but
finite effective bandwidth. Using polychromatic radiation always gives a negative deviation from
Beer’s law, but is minimized if the value of ε is essentially constant over the wavelength range
passed by the wavelength selector. In addition, deviations from Beer’s law are less serious if the
effective bandwidth from the source is less than one tenth of the natural bandwidth of the
absorbing species. When measurements must be made on a slope, linearity is improved by using
a narrower effective bandwidth.
Stray radiation is the second contribution to instrumental deviations from Beer’s law. Stray
radiation arises from imperfections within the wavelength selector that allows extraneous light to
“leak” into the instrument. Stray radiation adds an additional contribution, Pstray, to the radiant
power reaching the detector; thus

For small concentrations of analyte, Pstray is significantly smaller than P0 and PT, and the
absorbance is unaffected by the stray radiation. At higher concentrations of analyte, however,
Pstray is no longer significantly smaller than PT and the absorbance is smaller than expected. The
result is a negative deviation from Beer’s law.

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