INTRODUCTION TO

SPECTROCHEMICAL
METHODS
CEB 4032: ANALYTICAL CHEMISTRY

TS.DR. MOHD DZUL HAKIM WIRZAL

1
 Outline
▪ Electromagnetic Radiation
▪ Absorption Process
▪ Emission Process
▪ Qualitative Analysis
▪ Quantitative Analysis
▪ Analysis of Mixture
▪ Spectrometric Instrumentation

2
Learning Outcomes:
At the end of this chapter:

(1) Understand the properties of electromagnetic

radiation, absorption and emission processes in
spectrochemical methods.

(2) Make quantitative calculations, relating the amount of

radiation absorbed to the concentration of an
absorbing analyte.

(3) Describe the spectrometric instrumentation requires

for making measurements.

3
 INTRODUCTION
- Measurement based on light and other forms of
electromagnetic radiation are widely used throughout
analytical chemistry.

- The interactions of radiation and matter (absorption,

emission and fluorescence) are the subject of the
science call spectroscopy or spectrochemical
method.

- Spectrochemical methods are based on measuring

the amount of radiation produced or absorbed by
molecular or atomic species of interest.

4
- Can be classified according to the region of the
electromagnetic spectrum involved in the measurement.

- Regions that have been commonly used: X-ray, ultraviolet

(UV), visible, infrared (IR), microwave etc.

- Widely used tools for the elucidation of molecular

structure as well as quantitative and qualitative
determination of both inorganic and organic compounds.

- For example, the sample solution absorbs

electromagnetic radiation from an appropriate source and
the amount absorbed is related to the concentration of the
analyte in the solution.

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 Outline
▪ Electromagnetic Radiation
▪ Absorption Process
▪ Emission Process
▪ Qualitative Analysis
▪ Quantitative Analysis
▪ Analysis of Mixture
▪ Spectrometric Instrumentation

6
 Electromagnetic Radiation
▪ Electromagnetic radiation (EMR) → a form of
radiant energy that is propagated as a transverse
wave.

▪ General properties of EMR: i) wave and ii) particle

(discrete of energy) properties.

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8
 i) Characteristics of Wave

▪ Amplitude (A): Length of the electric vector at a maximum in the wave.

▪ Period (p): The time in seconds required for the passage successive
maxima or minima through a fixed point in space.

▪ Frequency (ѵ): The number of oscillations of the field that occur per
second and is equal to 1/p. (s-1 or Hz).

▪ Wavelength (λ): The linear distance between any two equivalent points
on successive waves (successive maxima or minima). (Å, nm, etc.).
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▪ Velocity (v):The relationship between the wavelength and
frequency (the speed of light)

frequency, s-1 or Hz

velocity, cm/s wavelength, cm

▪ In vacuum, the velocity of radiation is independent of wavelength

and it at its maximum. The velocity, c has been determined to be
2.99792 x 108 m/s.

▪ The above equation can be written as :

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▪ Wavenumber : reciprocal of the wavelength (cm-1).

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 ii) Characteristics of Particle
▪ When EMR is emitted or absorbed, a permanent
transfer of energy from the emitting object or to the
absorbing medium occurs.

▪ In this case, EMR is not a collection of wave but as a

stream of discrete particle → photons (the energy of a
unit of radiation).

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▪ Each photon carries energy, E, which is given by:

E = hѵ
frequency, s-1 or Hz

Plank’s constant = 6.626 x 10-34 J.s

▪ Energy is proportional to frequency, we can write:

▪ The shorter the wavelength or the greater the

frequency, the greater the energy.

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 Electromagnetic Spectrum
▪ encompasses an enormous range of wavelength and
frequencies. Spectroscopic methods are classified
according to the wavelengths or frequencies that are
important for analytical purpose.

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Common Spectroscopy Methods Based
on EMR

1 Å = 10-10 m = 10-8 cm
1 nm = 10-9 m = 10-7 cm
1 μm = 10-6 m = 10-4 cm
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16
 Example 1

In vacuum, what is the wavelength of light that

has a frequency of 5.00 x 1014 Hz? Into what
range of the light spectrum does it fall?

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 Solution

frequency, s-1 or Hz

velocity, cm/s wavelength, cm

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 Example 2:

i) By how many kilojoules per mole is the

energy of O2 increased when it absorbs
ultraviolet radiation with a wavelength of 147
nm?

ii) How much is the energy of CO2 increased in

kilojoules per mole when it absorbs infrared
radiation with a wavenumber of 2300 cm-1?

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Solution for (i):

(3.00 x 108 m/s)

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 Solution for (ii):
For CO2, the energy increase is

and

Given : wavenumber, 1/λ = 2300 cm-1

E = (6.626 x 10-34 J.s)(3.00 x 108 m/s)(2300 cm-1)(100 cm/m)

= 4.6 x 10-20 J/molecule x 6.022 x 1023 molecules/mol
= 28 kJ/mol

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 Exercise
Q1
Convert the wavelength 4000.0 Å into frequency
(Hz) and into wavenumbers (cm-1).

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Q2
What is the energy in kJ/mol possessed by a
radiation of wavenumber 1000.0 cm-1?

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Q3
Calculate the wavelength and energy in kJ/mol
associated with a signal at 220.0 MHz.

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 Outline
▪ Electromagnetic Radiation
▪ Absorption Process
▪ Emission Process
▪ Qualitative Analysis
▪ Quantitative Analysis
▪ Analysis of Mixture
▪ Spectrometric Instrumentation

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 Absorption of Radiation
▪ When radiation passes through a sample (a layer of
solid, liquid, or gas), certain frequencies may be
selectively removed by absorption.

▪ a process in which electromagnetic energy is

transferred to the atom, ions, or molecules composing
the sample.

▪ Absorption promotes these particles excited from

ground state (lower energy), to one or more higher
energy states.

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▪ The transition from one energy state to another, the
frequency or the wavelength of the radiation is
related to the energy difference between the states
by the equation:

Energy of the photon

E1-Eo = hѵ = hc/λ
Energy of the Energy of the
higher state lower state

▪ For absorption of radiation to occur, the energy of

the exciting photon must exactly match the energy
difference between the ground state and excited
state of the absorbing species.
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• These energy differences are unique for each
species, a study of the frequencies of absorbed
radiation provides a means of characterizing the
constituents of a samples of matter.

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Examples of UV absorption spectra

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 Atomic Absorption
▪ The passage of radiation through a medium that
consists of monoatomic particles results the
absorption of a few well defined frequency.

▪ The relative simplicity of such spectra is due to the

small number of possible energy states for the
absorbing particles.

▪ Excitation occur by an electronic process in which one

or more of the electrons of the atom are raised to a
higher energy level.

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Atomic Absorption

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 Molecular Absorption
▪ Absorption spectra for polyatomic molecules are
considerably more complex than atomic spectra.

▪ Because the number of energy states of molecules is

generally enormous when compared with the number of
energy states for isolated atoms.

▪ The energy E associated with the bands of a molecule is

made up of three components.

E = Eelectronic + Evibrational + Erotational

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• Eelectronic
- energy produced when the electrons of the molecule
raised to a higher energy level.

• Evibrational
- the total energy associated with the interatomic
vibrations that are present in molecular species.
- generally, a molecules has more vibrational energy
level than it does electronic levels.

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• Erotational
- is the energy caused by various rotational motions
within a molecule.
- the number of rotational states is much larger that the
vibrational stages.

The relative energy levels are in the order:

electronic>vibrational>rotational

Rotational → very low energy (low wavelength, microwave or

far-infrared region)
Vibrational →requires higher energies (near-infrared region)
Electronic → require still higher energies (visible and ultraviolet
regions)
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Energy Level Diagram

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 Outline
▪ Electromagnetic Radiation
▪ Absorption Process
▪ Emission Process
▪ Qualitative Analysis
▪ Quantitative Analysis
▪ Analysis of Mixture
▪ Spectrometric Instrumentation

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 Emission of Radiation
▪ Emission of radiation is occurred when excited
particle (atoms, ions, or molecules) relax to lower
energy levels by giving up their excess energy as
photons.

▪ Radiation from an excited source is characterized by

means of an emission spectrum.

X → X* →X + hѵ
Absorption

Emission

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Emission Process
Excitation

Emission

Emission Spectrum

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Emission Spectrum
▪ Three type of emission spectra:
1) Line spectrum
- made up of a series of sharp, well-defined peaks
caused by excitation of individual atoms.

2) Band spectrum
- consists of several groups of lines so closely
spaced that they are not completely resolved.

3) Continuum spectrum
- responsible for the increase in the background that is
evident above about 350 nm. The line and band
spectra are superimposed on this continuum.

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Example Emission Spectrum

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Energy-level diagrams for
(a) a atom showing the source of a line spectrum and
(b) a simple molecule showing the source of a band spectrum
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 Outline
▪ Electromagnetic Radiation
▪ Absorption Process
▪ Emission Process
▪ Qualitative Analysis
▪ Quantitative Analysis
▪ Analysis of Mixture
▪ Spectrometric Instrumentation

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 Qualitative Analysis
▪ Infrared spectroscopy is a very useful tool for obtaining
qualitative information about molecules.

▪ The most important used of infrared spectroscopy is the

identification and structure analysis.

Infrared Spectra
- The absorption peaks are much sharper than in the
ultraviolet or visible regions → easier to identify.

- Each molecule will have a complete absorption spectrum

unique to that molecule, and so a “fingerprint” of the
molecule is obtained.

- Mixtures of absorbing compounds → exhibit the combined

spectra of compounds.

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Examples: IR spectrum of compound

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Simple correlations of group vibrations to regions of infrared absorption

Typical functional groups that can be identified include alcohol, hydroxyl,

ester carbonyl, olefin and aromatic unsaturated hydrocarbon group.

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 Quantitative Analysis
▪ Quantitative absorption methods require two power
measurement: one before a beam (incident radiation)
has passed through the medium that contains the
analyte (Po) and the other after (P).

▪ Two terms, which are widely used in absorption

spectrometry related to the ratio of Po and P, are
transmittance and absorbance.

P<Po (absorption depends on population and path length)

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Transmittance

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Absorbance

A = -log T = log (1/T) =log (Po/P)

P/Po =0.1,

P/Po %T A
1 100 0
0.1 10 1
0.01 1 2
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49
Beer’s Law
▪ Basic for quantitative analyses by both atomic and
molecular absorption measurement.

▪ For monochromatic radiation, absorbance is directly

proportional to the path length b through the medium
and the concentration c of the absorbing species.

A = abc

a is a proportionality constant called absorptivity.

b is commonly expressed in centimeters (cm) and
c is usually given in unit grams per liter (g L-1).
Therefore, a has units of L g-1cm-1.
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When the concentration expressed in moles per
liter, the absorptivity is called molar absorptivity:

A = εbc

c is in moles per liter (mol/L or M) and ε has the

units L mol-1cm-1.

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 Example 3

Find the absorbance and transmittance of a

0.00240 M solution of a substance with a molar
absorptivity of 313 M-1 cm-1 in a cell with a 2.00
cm path length.

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 Solution
A =εbc
= (313 M-1 cm-1) (2.00cm) (0.00240 M) = 1.50

Transmittance:
A = -log (T)
T = 10-A
= 10-1.5
= 0.0316

3.16% of the incident light emerges from this

solution.

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 Example 4

A sample in a 1.0 cm cell is determined with a

spectrometer to transmit 80.0% light at a certain
wavelength. If the absorptivity of this substance
at this wavelength is 2.0, what is the
concentration of the substance?

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 Solution
The percent transmittance is 80 %, and so T = 0.80.

A = -log10(T) = abc

-log (0.80) = 2.0 cm-1g-1L x 1.0 cm x c

0.097= 2.0 g-1L x c

c = 0.097/2 = 0.048 g/L

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 Example 5

A solution containing 1.00 mg ion ( as the

thiocyanate complex) in 100.0 mL was observed
to transmit 70.0% of the incident light compared
to an appropriate blank.
(a) What is the absorbance of the solution at this
wavelength?
(b) What fraction of light would be transmitted by
a solution of iron four times as concentrated?

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 Solution

A = -log10(T)
= -log10(0.7)
= 0.155

0.620 = -log10(T), T= 0.240

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0.620 = -log10(T), T= 0.240

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 Deviation from Beer’s Law
▪ Beer’s law states that absorbance is proportional to
the concentration of the absorbing species.

▪ It applied to monochromatic radiation and it works

very well for dilute solution (≲ 0.01 M) of most
substances.

▪ Deviations occur as the result of (1) chemical, and (2)

instrumental factors.

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▪ Chemical deviations
→ from Beer’s law are caused by shifts in the position
of a chemical or physical equilibrium involving the
absorbing species.

▪ Instrumental deviations
→Unsatisfactory performance of an instrument may
be caused by fluctuations in the power-supply
voltage, an unstable light source, or a non-linear
response of the detector-amplifier system.

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 Exercise
Q1
Chloroaniline in a sample is determined as the
amine picrate. A 0.0265 g sample is reacted with
picric acid and diluted to 1.00 L. The solution
exhibits an absorbance of 0.368 in a 1.00 cm
cell. What is the % chloroaniline in the sample?
(Given: MW of Chloroaniline = 127.6 g/mol, ϵ =
1.25 x 104 cm-1mol-1 L).

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Q2
a. A 3.96 x 10-4 M solution of compound A exhibited
an absorbance of 0.624 at 238 nm in a 1.00 cm
cuvet; a blank solution containing only solvent
has an absorbance of 0.0290 at the same
wavelength. Find the molar absorptivity of
compound A.

b. A concentrated solution of compound A in the

same solvent was diluted from an initial volume
of 2.00 mL to a final volume of 25.0 mL and then
had an absorbance of 0.733. What is the
concentration of A in the concentrated solution?

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 Outline
▪ Electromagnetic Radiation
▪ Absorption Process
▪ Emission Process
▪ Qualitative Analysis
▪ Quantitative Analysis
▪ Analysis of Mixture
▪ Spectrometric Instrumentation

63
 Analysis of Mixture
▪ It is possible to make quantitative calculations when two
absorbing species in solution.

▪ It is apparent from Beer’s law that the total absorbance A,

at a given wavelength will be equal to the sum of the
absorbances of all absorbing species.

▪ For two absorbing species, if c is in g / L:

A=axbcx + aybcy

▪ If c is in mol/ L:
A=εxbcx + εybcy
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Pure substance, x

Mixture of x and y

Pure substance, y

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• From Figure 16.10 →two unknowns, x and y → two
measurements will have to be made.

• Technique: choose two wavelengths for measurement.

(1) occurring at the absorption maximum for x ( λ1 in
the figure).
(2) the other maximum for y (λ2 in the figure)

We can write:
A1 =Ax1 + Ay1 = εx1bcx + εy1bcy
A2 =Ax2 + Ay2 = εx2bcx + εy2bcy

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 A1 =Ax1 + Ay1 = εx1bcx + εy1bcy
A2 =Ax2 + Ay2 = εx2bcx + εy2bcy

• A1 and A2 are the absorbances for the mixture at

wavelength 1 and 2, respectively.

• Ax1 and Ay1 are the absorbances contributed by pure

substances x and y, respectively at wavelength 1.

• Ax2 and Ay2 are the absorbances contributed by pure

substances x and y, respectively at wavelength 2.

• εx1 and εy1 are the molar absorptivities contributed by

pure substances x and y, respectively at wavelength 1.

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• εx2 and εy2 are the molar absorptivities contributed
by pure substances x and y, respectively at
wavelength 2.

• These molar absorptivities (εx1, εy1,εx2 and εy2) are

determined by making absorbance
measurements on pure solution (known molar
concentrations) of pure substances x and y at
wavelength 1 and 2.

• cx and cy → two unknowns → calculated from the

solution of the two simultaneous equations
A1 =Ax1 + Ay1 = εx1bcx + εy1bcy
A2 =Ax2 + Ay2 = εx2bcx + εy2bcy
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 Example 6
The molar absorptivitives of compound X and Y were
measured with pure samples each

λ (nm) ε (M-1 cm-1)

X Y
272 16440 3870
327 3990 6420

A mixture of compounds X and Y in a 1 cm cell had an

absorbance of 0.957 at 272 nm and 0.559 at 327 nm.
Find the concentrations of X and Y in the mixture.

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 Solution
From Equations analysis of mixture:
A1 =Ax1 + Ay1 = εx1bcx + εy1bcy
A2 =Ax2 + Ay2 = εx2bcx + εy2bcy

A272 = εx,272b[X] + εy,272b[Y]

A327 = εx,327b[X] + εy,327b[Y]

Substitution: (b = 1 cm)
0.957 = 16440 [X] + 3870 [Y] (1)
0.559 = 3990 [X] + 6420 [Y] (2)

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Solve Equations (1) and (2) simultaneously:

0.559 = 3990 [(0.957-3870[Y])/16440] + 6420 [Y]

0.3267 = 5481 [Y]
[Y] = 5.96 x 10-5 M

Substitute [Y] into Equation (1) or (2),

[X] = 4.42 x 10-5 M

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 Example 7
Potassium dichromate and potassium permanganate have
overlapping absorption spectra. K2Cr2O7 has an absorption
maximum at 441 nm, and KMnO4 has a band at 545 nm. A mixture
is analyzed by measuring the absorbance at these two wavelengths
with the following results:

A441 = 0.405, A545 = 0.712

The absorbances of pure solution of K2Cr2O7 (1.00 x 10-3 M) and

KMnO4 (2.00 x 10-4 M), using the same cell gave the following
results:

ACr, 441 = 0.374, ACr,545 = 0.009, AMn,441=0.019, AMn, 545= 0.475.

Calculate the concentrations of dichromate and permanganate in

the sample solution.

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 Solution
The pathlength b is unknown precisely; but since the same cell is used
in all measurements, it is constant. We can calculate εb from the pure
substances measurement:

Mixture:
At A =441nm,
A441 = εCr,441b [Cr2O72-] + εMn,441b [MnO4-] ---------------(1)

At A = 545 nm,
A545 = εCr,545b [Cr2O72-] + εMn,545b [MnO4-] ----------------(2)

Pure substances:
At 440 nm,
0.374 = εCr,441b x 1.00 x 10-3, εCr,441b = 374
0.019 = εMn,441b x 2.00 x 10-4, εMn,441b = 95

At 545 nm,
0.009 = εCr,545b x 1.00 x 10-3, εCr,545b = 9
0.475 = εMn,545b x 2.00 x 10-4, εMn,545b = 2.38 x 103

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Substitution of ε values into Equations (1) and (2) :

0.405 = 374[Cr2O72-] + 95[MnO4-]

0.712 = 9[Cr2O72-] + 2.38 x 103[MnO4-]

Solving simultaneously,

[Cr2O72-] = 1.01 x 10-3 M

[MnO4-] = 2.95 x 10-4 M

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 Example 8
Analysis of an unknown mixture of H2O2 complexes of Ti(IV) and V(V) in H2SO4
Solution.

Visible spectra of standard solution: 1.32 mM Ti(IV), 1.89 mM V(V),

and an unknown mixture containing both ions. All solution contain 0.5 wt%
H2O2 and ~ 0.01 M H2SO4.
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 Absorbance Data

Wavelength AXs Titanium AYs Vanadium Am

(nm) standard standard Mixture

390 0.895 0.326 0.651

430 0.884 0.497 0.743
450 0.694 0.528 0.665
470 0.481 0.512 0.547
510 0.173 0.374 0.314

Determine the concentration of the unknown mixture.

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The absorbance of the mixture (Am) at any chosen
wavelength is

Am = εxb[X] + εyb[Y] Mixture

Where X and Y refer to Ti(IV) and V(V), respectively. If a

standard solution of species X with concentration [X]s is
prepared, its absorbance will be

Axs = εxb[X]s

Similarly, for species Y, [Y]s Pure Component

Ays = εyb[Y]s

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Solving the above equations for εx and εy,

How???

Slope Intercept

Given in the question 78

Wavelength AXs AYs Am Am/AXs AYs/AXs
(nm) Titanium Vanadium Mixture
standard standard
390 0.895 0.326 0.651 0.727 0.364
430 0.884 0.497 0.743 0.840 0.562
450 0.694 0.528 0.665 0.958 0.760
470 0.481 0.512 0.547 1.137 1.064
510 0.173 0.374 0.314 1.815 2.161

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The least-square straight line through the point is:

Therefore,

Slope = 0.607 = [Y]/[Y]s

→ [Y] = [V(V)] = 0.607 x 1.89 mM = 1.15 mM

Intercept = 0.499 = [X]/[X]s

→ [X] = [Ti(IV)] = 0.499 x 1.32 mM = 0.659 mM

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 Outline

▪ Electromagnetic Radiation
▪ Absorption Process
▪ Emission Process
▪ Qualitative Analysis
▪ Quantitative Analysis
▪ Analysis of Mixture
▪ Spectrometric Instrumentation

81
 Spectrometric Instrumentation
▪ Spectrometer or Spectrophotometer → an
instrument/device to measure absorbance of light that
resolve polychromatic radiation into different
wavelength.

▪ Consists of
(1) a source of continuous radiation over the
wavelength of interest.
(2) a monochromator for selecting a narrow band of
wavelengths from the source spectrum.
(3) sample cell for holding sample.
(4) a detector or transducer → for converting radiant
energy into electrical energy.
(5) a device to read out the response of the detector.
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Block diagram of spectrometer/spectrophotometer
 (1) Sources of Light
▪ It’s provides radiation of range of wavelengths.

Tungsten lamp:
- Produces visible and near-infrared radiation in the range
from 320-2500 nm.

Deuterium Arc Lamp:

- Produces UV radiation in the range from 200 - 400 nm.

84
 Source of light: Lamps

Deuterium Lamp Tungsten Lamp

85
 (2)Monochromators

▪ Disperse light into its component wavelength and

selects as a narrow band of wavelengths to pass on
to the sample.

▪ Consists of (1)entrance and exit slits, (2) mirrors, and

a (3) grating to disperse the light.

▪ Two basic types of dispersing elements (1) prism and

(2) diffraction grating
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Diffraction Grating:
linear dispersion, independent
of wavelength , but the
intensity varies with
wavelength. Eg.: aluminium

Prism: non-linear
dispersion, good at short
wavelength, poor at long
wavelength. Eg.: glass or
lenses.
87
 (3)Sample Cell

▪ The cell holding the sample → transparent in the

wavelength region being measured.

▪ For visible and ultraviolet spectrophotometers, a liquid

sample is usually contained in a cell called a cuvet → flat,
fused silica faces. Most common used cuvet : 1.00 cm
pathlength.

▪ Glass → for visible, not for ultraviolet because it absorbs

UV radiation.
▪ For IR measurement, 1 wt% mixture of solid sample with
KBr → grounded to fine powder and pressed into
translucent pellet for mounting and measurement.
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Cuvet for UV/Vis
Measurement

89
FT-IR Sample Preparation- KBr + Sample Pellet

Pelletizer

Agitate mortar
Sample holder

Die

90
 (4) Detectors

▪ A detector produces an electric signal when it is

struck by photons.

Examples UV-Vis detector:

(1) Phototube or photocell

(2) Photomultiplier (PM) tube
(3) Photodiaode array

91
Phototubes Photomultiplier

Photodiaode array
92
IR detectors:
- Detectors used in the UV-Vis regions do not work in
the IR region.

- IR photon do not have sufficient energy to generate a

signal in either kind of detector.

- IR radiation possesses the property of heat, and heat

detectors that transduce heat into an electrical signal
can be used.

Examples IR detector:
(1) Thermocouple
(2) Balometers
(3) Thermistors
93
Thermocouples

Balometers and Thermistors

94
 MEASUREMENT OF
ABSORBANCE AND
TRANSMITTANCE

1) Single-beam spectrophotometer
2) Double-beam spectrophotometer

95
 Single- beam Spectrophotometer

General operation:

1) The radiant power of light passing through a reference cuvet

containing pure solvent is defined as Po.

2) The cuvet is then removed and replaced by sample cuvet,

the radiant power → P.

3) Knowing P and Po→ allows T or A to be determined.

4) Any radiation not absorbed by the sample falls on the

detector→ intensity converted to an electrical signal.
96
 Single- beam Spectrophotometer

▪ One beam of light.

▪ Inconvenient because the sample and

reference must be placed alternately in the
beam.

▪ For measurements at multiple wavelength,

the reference must be run at each
wavelength.

▪ Poorly suited to measuring absorbance as a

function of time.

97
Double - beam Spectrophotometer

98
 Double-beam Spectrophotometer
▪ Has two light path → sample and blank reference.

▪ Through a rotating mirror, the light beam alternately

passes through the reference cell and the sample cell,
from each, to the detector.

▪ Automatically vary the wavelength and record the

absorbance as a function of wavelength.

99
END OF CHAPTER

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