Annachapter 6 Spectros

Analytical Techniques 6.
CHAPTER-6
ANALYTICAL TECHNIQUES
6.1 INTRODUCTION
To Recapitulate the Wave parameters

The electromagnetic radiation can be described in a number of
properties of parameters.
Wavelength ()
It is the distance between the two adjacent crests (C-C) or
troughs (T-T) in a particular wave. It is denoted by the letter
( lam da)
C  C
Units used for wave length are :
Angstrom(A0) = 10-10m = 10-8 cms
Micrometer (m) = 10-6m
Nano meter (nm) = 10-9m 
T T
Frequency ()
Frequency may be defined as the number of waves which
can pass through a point in one second (i.e. number of vibrations
in unit time) of an electromagnetic radiation.
Frequency () = Velocity of light / Wavelength
 = c/
Velocity of light c = 3  1010 cm/sec
Wavelength  in cm
6.2 Engineering Chemistry-II
The unit for frequency is cycles per second or in (Hz).


Wave number ( )
The number of waves per unit length is known as wave number
and is expressed as the reciprocal of wave length.
 1
 

Wave number is expressed as the waves per cm (cm-1). This unit

is also called Kayser (K).
The wave length, the frequency and the wave number are
related as follows :
1  
= =
 c
6.2 ELECTROMAGNETIC RADIATION
Electromagnetic radiation is a type of energy that is

transmitted through space at enormous velocities. It takes many
forms, the most easily recognizable being light and radiant heat.
Less obvious manifestations include X-ray, Ultraviolet,
microwave and radio radiations.
The electromagnetic radiation is found to have dual nature

- the particle nature (photons) and wave nature.
The principles and laws that govern the absorption of

radiation apply to all wavelengths form X-ray to radiofrequency.
The absorption measurements involve the determination of the
reduction in power suffered by a beam of radiation as it passes
through an absorbing medium of known dimensions.
Analytical Techniques 6.3
Table 6.1 - The regions of electromagnetic spectrum
Type of transition resulting Frequency Type of Wavelength

from adsorption Hz spectro
metry
Change of Inner electron 3x 1016 – X-ray 10nm –
electronic 3x 1018 100pm
distribution Outer electron 3x 1014 – Visible 1micro m –
3x 1016 - UV 10 nm
Change of Molecular 3x 1012 – IR 100microm
configurat-on vibrations 3x 1014 – 1microm
Change of Molecular 3x 1010 – Micro 1cm –
orientations rotations 3x 1012 wave 1 micro m
Change of spin Spin 3x108 – ESR 100cm –
orientations 3x 1010 1 cm
Spin 3x106 – NMR 10 m –
orientations 3x 108 Radio 100cm
waves
When the monochromatic radiation passes through a

sample containing an absorbing species, the radiant power of a
beam is progressively decreased as more energy is absorbed by
the particles of that species. The decrease in power is expressed
by
1) Lambert’s law
2) Beer’s law (Or) Beer - Lambert’s law
Lambert’s law
Lambert’s law states that “when a beam of
monochromatic radiation is passed through a homogeneous
absorbing medium, the rate of decrease of intensity of the
radiation ‘dI’ with thickness of absorbing medium ‘dx’ is
proportional to the intensity of the incident radiation ‘I’ ”.
 dI
i.e,  kI ……… (1)
dx
On integrating the equation (1), between the limits I = I0 at x = 0

and I = I at x = x, we get
I x
dI

0
I
   k dx
0
……… (2)
where, k = absorption coefficient
I
i.e., ln  k x
I0
The above equation is known as Lambert’s law.
Beer’s law (or) Beer-Lambert’s ’s law
When the monochromatic radiation passes through a

sample containing an absorbing species, the radiant power of a
beam is progressively decreased as more energy is absorbed by
the particles of that species. The decrease in power depends upon
the concentration of the absorber and the length of the path
traversed by the beam. The relationship is expressed by Beer’s
law.
Let I0 be the radiant power of a beam incident upon a section of

solution that contains c moles of an absorbing substance per liter.
Let I be the power of the beam after it has traversed x centimeters
of the solution. As a consequence of absorption, I will be smaller
than I0. Beer’s law relates these quantities as
 I 
log     cx  A
 I0 
In this equation is called as the molar absorptivity. The
logarithm (to base 10) of the ratio between the incident and the
transmitted power is called the absorbance of the solution.
The absorbance increases directly with the concentration

of the absorbing species and the path length traversed by the
beam.
Thus Beer-Lambert’s law states that, “when a beam of

monochromatic radiation is passed through a solution of an
absorbing substance, the rate of decrease of intensity of
radiation ‘dI’ with thickness of the absorbing solution ‘dx’ is
proportional to the intensity of the incident radiation ‘I’ as
well as the concentration of the solution ‘C’.
It is expressed as
 dI
 kIC …… (3)
dx
where , k = molar absorption coefficient.
On integrating the equation (3) between limits I = I0 at x = 0 and

I = I at x = x , we get,
I x
dI

I0
I
   k C dx
0
I
ln  kC x
I0
I
(or) 2.303 log I   k C x
0
I k
log 0  Cx
I 2.303
(or) A  Cx ……. (4)

k
Where ,   = molar absorptivity coefficient
2.303
I0
log  A  Absorbance (or) optical density
I
Equation (4) is called Beer-Lambert’s law.
Thus, the absorbance (A) is directly proportional to molar

concentration ‘C’ and thickness (or) path length (x).
Application of Beer- lamberts law
Determination of Unknown concentration of a solution

Beer –Lambert’s law can be used to determine the
unknown concentration of a solution
A standard solution of known concentration Cs is first
taken and it’s absorbance As is measured .
Then according to Beer-Lambert’s law
As   C s x
As
  x ………. (1)
Cs
Let Cu be the unknown concentration of the solution which is to

be determined.
The absorbance Au of the given solution is measured.
We know that
Au   C u x
Au
   x ……..….(2)
Cu
From equation (1) and (2), we get

As Au

Cs Cu
Au
 Cu   Cs ………. (3)
As
We already know the concentration of standard solution
Cs and the absorbance Au and As were experimentally measured
and hence the unknown concentration Cu can be calculated from
the equation (3).
Limitations of Beer-Lambert’s law :
This law can be used only for

dilute solution.
monochromatic radiation.
the system maintained at constant temperature.
Solved Problems :
1) The transmittance of a solution is 30%. Determine the

absorbance (or) optical density of the solution.
Solution
Transmittance T = 0.30
Absorbance is
1
A = log (or)  log T
T
= – log 0.30
= 0.5229
Example 2: The solution of concentration of 2.5 10-4M has the

percentage absorption of 25% in a 1 cm cell. Determine
i) the absorbance (A)and ii) the molar absorption coefficient ().
Solution :
Given : % T = 25% (or) 0.25
C = 2.5 10-4M ; x = 1
i) Absorbance :
Absorbance, A   log T
= –log 0.25
= 0.6021
ii) The molar absorption coefficient

A
 
Cx
= 0.6021 / 2.5 x 10-4
= 2408 mol 1cm 1 .
Example 3: The absorbance of a solution 1.2  10-4 M. is found

to be 0.23 in a path length of 3.0 cm. Determine the molar
absorption of the solution.
Solution :
Given : A = 0.23 ; C = 1.2  10-4 ; x = 3.0 cm
We know that,
A
Molar absorptivity,  
Cx
0.23
=
1.2  10  4  3.0
= 638.89 dm3 mol-1 cm-1.
Example 4: A solution shows a transmittance of 20% when taken

in a cell of 2.5 cm thickness. Calculate it’s concentration if the
molar absorption coefficient is 12,000 dm3 mol-1 cm-1
(Anna Univ, Jan’2005)
Solution :
Given : T = 20% (or) 0.20 ; x = 2.5 ;
  12,000 dm3 mol-1 cm-1
A   log T   log 0.20
 0.699
We know that,
A   Cx
A 0.699
 C  = 12,000  2.5
 x
= 2.33  10 5 mol 1 dm 3 .
Example 5 : A solution of thickness 2 cm transmits 40%

incident light. Calculate the concentration of the solution
given that  = 6000 dm3 mol-1 cm-1.
(Anna univ June 2005)
Solution
Given : T = 40% = 0.40;  = 6000 dm3 mol-1 cm-1.
x = 2.0 cm;
Absorbance, A   log T
  log 0.40
 0.3980
we have,
A   Cx
A 0.3980
C  =
 x 6000  2
= 3.316 10–5 mol dm–3
Example 6 :
A monochromatic radiation is incident on a solution of 0.06 M

concentration of an absorbing substance. The intensity of the
radiation is reduced to one-fourth of the initial value after passing
through 8 cm length of the solution. Calculate the molar
extinction co-efficient of the substance. (Anna Univ May 2004)
Solution:
I 1
Given : C = 0.06 M ; x = 8 cm I  4 ,   ?
0
According to Beer-Lambert law,
I
log    Cx
I0
 log I I 0  log 1 4 
   
Cx 0.06  8
 ( 0.6020)

0.06  8
 = 1.254 dm3 mol-1 cm-1
________________________________________________
6.3 INTRODUCTION TO SPECTROSCOPY
All chemical species interact with the electromagnetic

radiation and in doing so, diminish the intensity or the power of
the radiant beam. Absorption spectroscopy is based upon the
measurement of this decrease in power (or attenuation) of the
radiation brought about by the analyte.
It is convenient to characterize absorptiometric methods

according to the type of electromagnetic radiation employed.
They are X-rays, Ultraviolet, Visible, Infrared, microwave and
radio-frequency radiation.
A white source (electromagnetic radiation) is focused on

to a narrow slit and then made to pass through the sample. The
light is absorbed by the sample and this is separated into its
constituent frequencies by a filter and then sent to the recorder.
At this point it may be helpful to consider what happens to the
energy absorbed in the above process.
A given sample continues to absorb energy and show an

absorption spectrum as long as we irradiate it. i.e. a finite number
of molecules appear to be capable of absorbing an infinite
amount of energy. Plainly the molecules should get rid of the
absorbed energy.
A possible mechanism for this is by thermal collision. An

energized molecule collides with its neighbors and gradually
loses its excess energy to them as kinetic energy. The sample as
whole becomes warm.
Another mechanism is that the energy gained from

radiation is lost as radiation once more. A molecule in the ground
state absorbs energy at frequency and its energy is raised an
amount E = hv above the ground state. It is thus in an excited,
unstable condition, but by emitting radiation of frequency v
again, it can revert to the ground state.
The spectrum obtained could be either due to the excess

radiation which is not absorbed or due to the emitted radiation.
The study of spectroscopy can be carried out under the

following two heads :
Atomic Spectroscopy
Interaction of electromagnetic radiation with atoms which are
most commonly in their lowest energy state (ground state)
resulting in electronic transitions is called atomic spectroscopy.
The spectrum obtained is a line spectrum.
Molecular spectroscopy
Interaction of electromagnetic radiation with molecules is
called Molecular spectroscopy. This may result in transitions
between rotational, vibrational energy levels and electronic
transitions. The spectrum obtained is a complicated spectrum.
Differences between Atomic and Molecular spectra

Atomic spectra Molecular spectra
1 It is produced due to It is obtained from the
interaction of atoms interaction of molecules with
with electromagnetic electromagnetic radiation.
radiation
2 Atomic spectra are Molecular spectra are
Line spectra. complicated spectra.
3 It is obtained due to It is produced due to
electronic transition vibrational, rotational and
in an element electronic transition in a
molecule.
Absorption spectroscopy
If electromagnetic radiations of certain wavelength range
are passed through the substance under analysis, radiations of
certain wavelengths are absorbed by the substance. The study of
this is called the absorption spectroscopy. The wavelength
absorbed characterizes some specific functional group of the
compound or the compound itself.
Higher energy level
h
Ground state
Absorption
Higher energy level
h
Ground state
Emission
Emission spectroscopy
If electromagnetic radiation is passed through a substance
or thermal energy is given to the substance under analysis, the
energy is absorbed by the atom. The electrons in the ground state
get excited to higher energy metastable states. These excited
electrons are short lived. So they emit energy to return to the
stable state. The study of this is called the emission spectroscopy.
The spectrum obtained is called the emission spectrum.
Fluorescence
The electron in the excited level return to it’s ground state
either directly or in steps with the emission of certain amount of
energy. When this emission of light is instantaneous the
phenomenon is known as fluorescence
Phosphorescence
When the electron in the excited level return to it’s
ground state with the emission of light after some time lag, it is
known as phosphorescence
Photochemical reaction
When the absorbed energy is stored by the atom or
molecule and used in producing some chemical reaction, the
resulting chemical reaction is called photochemical reaction.
6.4 CALORIMETRY
Introduction
Colorimetry is concerned with the visible region (400-
750nm) of the spectrum. The instrument, used for measuring
absorption of radiant energy in the visible region from the
substances is colorimeter.
Preparation of the coloured solution :

Although some desired constituents are self coloured, it is
usually necessary to develop a colour by the addition of one or
more colour forming reagents. Chemical methods used to prepare
suitable coloured solutions are usually called chromogenic
reactions. The colour forming reagents are known as
chromogenic reagents.
Example : The complexing agent ammonium hydroxide is added

to get blue coloured solution and to estimate the cuprous ions
Instrumentation for colorimetry
Principle
Colorimetry is concerned with the determination of the
concentration of a substance by measurement of the relative
absorption of light in the visible region with respect to a known
concentration of substance. The intensity of the colour can be
easily measured by using a photoelectric colorimeter, from which
the concentration of coloured solution can be obtained by Beer –
Lamberts law.
I
A   log T  log 0   Cx
It
Current  Light transmitted  (1/ Concentration of solution)
Working
In a colorimeter, a beam of light is directed through a
transparent cell containing a solution of the compound being
analysed. The compound absorbs light at a particular wavelength.
The unabsorbed radiation (transmitted light) is passed through the
detector where the light signals are converted into the electrical
signals which are recorded. The output graph obtained is a plot of
the absorbance of the light versus concentration.
A = log(I0/I)
The amount of light transmitted depends on the depth of colour

of the test solution.
Components
Radiation sources
Tungsten – filament lamp
Function:
Provides the wavelength range of the visible region
(400-750nm).
Sample chamber
Cuvet made of fused glass
Function:
A transparent cell made of fused glass holds the test
sample
Slits - Function:
Entrance slits: It provide a narrow source of the light.
Exit slit: It selects a narrow band of dispersed spectrum and

sends it to the detector.
Filter
Monochromator
Function:
Allows the light of the required wavelength to pass
through, but absorbs the light of other wavelengths.
Detector
Photosensitive devices are used to detect the radiations.
Function:
Converts the radiation signals received from the sample
into the electrical signals.
These detectors are capable of producing current, which is
directly proportional to the intensity of the radiation received.
Thus the radiant energy transmitted through the sample is
measured.
Recorder
Digital display
Function:
It displays the amount of radiation absorbed by the
sample
COLORIMETRY - Instrumentation Block diagram
LIGHT FILTER SAMPLE

SOURCE DETECTOR
CELL
AMPLIFIER
RECORDER
MATERIALS USED IN COLORIMETRY
LIGHT SOURCE FILTER SAMPLE CELL

Tungsten filament Monochromator Cuvet made of
lamp fused glass
DETECTOR
Photo multiplier
tube
AMPLIFIER
RECORDER
Digital display
FUNCTIONS OF THE PARTS USED IN COLORIMETRY

LIGHT FILTER SAMPLE CELL
SOURCE Selects the Sample is kept in
Provides the required this
source of radiation wavelength
DETECTOR
Converts the light
signals into electrical
signals
AMPLIFIER
RECORDER
Displays the result
Estimation of Concentration of a solution

Colorimetry technique may be used to determine the
concentration of a substance from the coloured solution.
Estimation of copper sulphate

A series of known concentration of standard copper
sulphate solutions are prepared. To these definite volume of
NH4OH is added. These are the blue coloured solutions. The
colorimeter is set at zero absorbance using a blank solution, with
a proper filter. Now absorbance of each standard solution is
measured using the same filter. A graph is plotted between
absorbance vs concentration. This plot is called calibration curve
and will be the straight line passing through origin. This is
according to Beer-Lambert’s law.
Absorbance, A =  C x
Since, the path length (x) is fixed for a given cell, the absorbance
(A) is directly proportional to concentration (C). Similarly, to the
test solution same amount of NH4OH is added and the absorbance
of the test solution (unknown copper sulphate) is measured using
the same colorimeter. From the calibration curve, the
concentration of the unknown copper sulphate solution can be
evaluated.
Absorbance of
Unknown solution 

Absorbance 
 Concentration of
unknown solution

Concentration
Calibration curve
Estimation of Fe3+ (ferric) iron
Fe3+ is a colourless solution. So to the ferric solution,

KCNS (or) NH4CNS is added to give a blood red coloured
complex.
Fe3+ + 6NH4CNS  [Fe(CNS)6]3- + 6NH4+

(Blood red coloured complex)
A series of standard solution of thio cyanate complex of

Fe3+ (ferric ammonium sulphate) are prepared. The colorimeter is
set at zero absorbance using a blank solution, with a proper filter.
Now absorbance of each standard solution is measured using the
same filter. A straight line calibration curve is obtained by
plotting between absorbance vs concentration. This is according
to Beer-Lambert’s law.
Absorbance, A =  C x
Since, the path length (x) is fixed for a given cell, the
absorbance (A) is directly proportional to concentration (C).
Similarly, to the Fe3+ test solution, the same amount of thio
cyanate is added and the absorbance of the test solution is
measured using the same colorimeter. From the calibration curve,
the concentration of the unknown Fe3+ solution can be evaluated.
Absorbance of
Unknown solution 

Absorbance 
 Concentration of
unknown solution

Concentration
Calibration curve
Applications of colorimetry
 To determine the molar compositions of complexes.
 To measure the dissociation constants of an acid-base
indicator.
 To measure the molecular weight of a compound.
 To determine the instability constants of complexes.
 To determine the structure of inorganic compounds and
complexes.
6.5 ULTRA VIOLET SPECTROSCOPY
Introduction
UV consists of broad absorption bands which have greater
utility for quantitative analysis. When polychromatic UV and
Visible radiation is passed through a medium containing
particles, they interact with the species and electronic transitions
occur by the excitation of electrons from the ground state to the
excited state. UV and visible radiation have sufficient energy to
ensure transitions of the outermost or bonding electrons only.
Singlet set: On excitation the spins of the electrons in the

molecular orbitals are paired.
Triplet set: On excitation the spins of electrons in the
molecular orbital is unpaired or anti parallel
According to molecular orbital theory, the interaction of atomic

orbitals leads to the formation of bonding and antibonding
molecular orbitals. They may be classified as
Sigma (): The electron density is concentrated along the
internuclear axis.
Pi () type: The electron density is concentrated above and

below the internuclear axis.
Chromophores: The part of the molecule having n or 

electrons which is essentially for
absorption,.
Example C=C, C=O, N=N
Auxochromes: Polar groups having lone pair of electrons.

Example –OH, OR, -NH2, -SH etc
Instrumentation for UV and Visible spectroscopy
Principle
When UV-Visible radiations (wavelength range – 200-
800nm) are absorbed by atom / molecules, the transition of
valence electrons from lower electronic energy level to higher
electronic energy level occurs. The amount of energy required for
the transition depends on the difference in energy levels of
ground state and excited state.
E1 – E0 = h.
When excitation occurs, an electron from one of the filled ,  or

n molecular orbitals get excited to the vacant * or *
(antibonding) molecular orbitals. Corresponding to possible
excitations, there are various absorptions.
  *  * n  *
 *   * n  *
 electrons require higher energy for excitation to * or * than

the excitation of  electrons which in turn is greater than n
electrons.
The order of decreasing energy for the transition is
* > * = * > n* = * > n *

Working
In a UV-Visible spectrometer, a beam of light is split into

two halves. One half of the beam (the sample beam) is directed
through a transparent cell containing a solution of the compound
being analysed and one-half (the reference beam) is directed
through an identical cell that contains the solvent or blank
solution. The intensities of the two beams at each wavelength of
the region is compared. If the compound absorbs light at a
particular wavelength, then intensity of the sample beam will be
less than that of the reference beam.
The motor light attenuator drives an optical edge into the

reference beam until the detector receives light of equal intensity
from sample and reference beam. The detector converts UV
radiation into electrical signals and sends it to recorder. The
output graph obtained is a plot of the wavelength of the entire
region versus the absorbance of the light at each wavelength.
 I0 
A = log 
 I 
The graph obtained is called as absorption spectrum
Components :
The modern ultraviolet-visible spectrometers consist of
light source, sample chamber, Motor light attenuator, filter,
detector, amplifier and the recording devices.
Source of light :
 >375 nm : Tungsten Filament lamp
 < 375 nm : Hydrogen discharge lamp
Deuterium discharge lamp
Xenon discharge lamp
Mercury arc
Function
Provides the wavelength region corresponding to UV
radiation.
Tungsten filament lamp is particularly rich in red
radiations,
Sample chamber (sample cell and reference cell)

 Silica cell of path length 1cm
Function
Used to hold the sample and reference solutions. Silica
cells are transparent throughout the region
Chopper
 Circular disc with alternate quadrants removed.
Function
Rotates and transmits sample and reference beam
alternatively to grating
Filters :
 Monochromators - It consists of an entrance slit, a
dispersing element (prism or grating) and an exit slit.
 Grating made of quartz (OR) fused silica,
Function
Sends individual required frequencies to detector
Detectors :
 Barrier layer cell
 Photomultiplier tube
 Photocell
Function
The detector converts the radiation signals into
current. The current is directly proportional to the
concentration of the solution.
Motor light attenuator

Function
Drives an optical edge into the reference beam until the
detector receives light of equal intensity from sample and
reference beam.
Amplifier and Recording system

 Pen/ digital recorder
Function
The output graph which is a plot of the wavelength of the
entire region versus the absorbance of the light at each
wavelength is obtained.
ULTRA VIOLET SPECTROSCOPY :
Instrumentation Block diagram
LIGHT CHOPPER
SOURCE
REFERENCE
FILTER
DETECTOR
MOTOR LIGHT AMPLIFIER

ATTENUATOR
RECORDER
Applications of UV Spectroscopy
Ultraviolet spectroscopy has been mainly applied for the
detection of functional groups (chromophore), the extent of
conjugation, detection of polynuclear compounds by comparison
etc. Some important applications of UV spectroscopy are as
follows :
1. Detection of functional groups :

This technique is applied to detect the presence or
absence of the chromophore. The options of a band at a particular
wave length may be regarded as an evidence for the absence of a
particular group in the compound.
2. Extent of conjugation :
The extent of conjugation in polyenes can be estimated.
3. Distinction in conjugated and non-conjugated

compounds :
It also distinguishes between a conjugated and a non-
conjugated compound. The following isomers can be readily
distinguished since one is conjugated and the other is not.
The forbidden n* band for the carbonyl group in the

conjugated compound will appear at longer wave length
compared to that for the non conjugated compound.
The alkyl substitution in an alkene causes a bathochromic

shift. The technique is not much useful for the identification of
individual alkenes.
4. Identification of an unknown compound

An unknown compound can be identified by comparing
it’s spectrum with the known spectra. If the two spectra coincide,
the two compounds must be identical. If the two spectra do not
coincide, then the expected structure is different from the known

compound.
5. Examination of Polynuclear hydrocarbons

Benzene and polynuclear hydrocarbons have
characteristic spectra in the ultra-violet and visible region. Thus
the identification of the polynuclear hydrocarbons can be made
by comparison with the spectra of known polynuclear
compounds. The presence of substituents on the ring, generally,
shifts the absorption maximum to longer wave length.
6. Detection of impurities:
UV absorption spectroscopy is used for detecting impurities
in organic compounds, because
i) Bands due to impurities are very intense.
ii) Saturated compounds have little absorption band and
unsaturated compounds have strong absorption band.
7. Elucidation of the structure of Vitamins A and K.
8. Identification of a compound in different solvents :

Sometimes, the structure of the compound changes with
the change in the solvent which could be identified
9. Quantitative analysis
Determination of substances :
UV absorption spectroscopy is used for the quantitative
determination of compounds, which absorbs UV.
First, absorbance (optical densities) of the different
solutions of known concentration are measured. Beer’s law is
used for this determination.
I0
A   log T  log   Cx
It
where,
 = molar extinction coefficient
C = concentration
x = length of the cell
Then the graph is plotted between absorbance vs concentration.
Thus we get the calibration curve in the form of a straight line.
From the graph, the concentration of unknown substance is found
out.
10. Determination of molecular weight
11. Determination of calcium in blood serum

Calcium in the blood can be determined by converting the
‘Ca’ present in 1 ml of the serum as it’s oxalate and redissolving
it in H2SO4 and treating it with dilute ceric sulphate solution. The
absorption of the solution is measured at 315 nm. Thus the
amount of Ca in the blood serum can be determined.
12. Determination of configurations of Geometrical

Isomers :
The results of absorption show that cis-alkenes absorb at
different wave-lengths as compared to their corresponding trans-
isomers.
6.6 INFRARED SPECTROSCOPY
Introduction
The atoms in a molecule do not remain in fixed relative
positions but vibrate about some mean position. This fact is used
in the IR spectroscopy. The infrared region in the electromagnetic
spectrum extends from the red end of the visible spectrum to the
microwave region. The region includes radiation at wavelengths
between 0.7 and 500 —m or, in wave numbers, between 14000
and 20 cm-1. The spectral region is split into three regions
Near IR (12,500 - 4000 cm-1)

The absorption bands resulting from harmonic overtones

of fundamental bands and combination bands are present. This
region is used for quantitative work.
Mid IR (4000 -650 cm-1)
Many useful correlations could be obtained from this
region. This region is divided into two regions - the group
frequency region 4000-1300 cm-1 and finger print region in
1300 -650 cm-1.
In the group frequency region the principal absorption
bands may be assigned to vibration units consisting of only two
atoms of a molecule i.e. to units which are more or less
dependent on the functional group.
In the finger print region are the single bond stretching

frequencies and bending vibrations of polyatomic systems which
involve motions of bands linking a substituent group to the
remainder of the molecule.
Far IR (667-10 cm-1)

Correlates the bending vibrations of C, N, O and F with
atoms of heavier mass. This is well suited for studying
organometallic and inorganic compounds whose atoms are heavy
and whose bonds are inclined to be weak.
Number of Fundamental vibrations

Consider a molecule containing N-atoms. The position of
each atom is referred to by specifying three coordinates (the x, y,
and z-coordinates). Thus the total number of coordinate values is
3N. Thus the molecule has 3N degrees of freedom.
The molecule is free to move in three dimensional space
without change of shape. i.e. there can be translational movement
which uses three degrees of freedom.
Also the molecule can possess the rotational movement.
The rotational movement of non linear molecule uses three
degrees of freedom whereas the linear molecule uses only two
degrees of freedom as there is no rotation in the bond axis.
Thus the number of fundamental vibrations for

Non linear molecule is 3N-6
Linear molecule is 3N-5
Instrumentation for Infra red spectroscopy
Principle
The IR spectra are given by diatomic molecules with
permanent dipole moments, i.e. heteronuclear atomic molecules
and polyatomic molecules. When IR radiations (wavelength
range –14000 – 20cm-1) are absorbed by atom / molecules, the
atoms/ molecules vibrate and undergo transition between
vibrational levels. The transitions in the vibrational energy levels
are also accompanied by transitions in the rotational energy
levels. (So IR is also called as Rotational – Vibrational spectra).
These results in the absorption bands that is characteristic to the
atoms / molecules.
The different vibrations are
Stretching vibration
The distance between two atoms changes, but the bond
angle is not altered.
Symmetric stretching
Either the movement of atoms occurs towards one
another or away from one another
Asymmetric stretching
One atom approaches the central atom while
the other departs from it .
Bending vibrations
The bond angle changes and the bond distance do not
change.
Scissoring (in – plane bending) -

Movement of atoms is towards each other
Rocking (in – plane bending) -

Movement of atoms is in same direction
Wagging (out of plane bending)

+ +
Atoms move up and down the plane with
respect to the central atom
Twisting (out of plane bending).

- +
One of the atoms move up and other moves down
the plane with respect to central atom
Working
In a IR spectrometer, a beam of light is split into two
halves. One half of the beam (the sample beam) is directed
through a transparent cell containing the compound being
analysed and one-half (the reference beam) is directed through an
identical cell that contains the solvent or blank. The intensities of
the two beams at each wavelength of the region is compared. As
the compound absorbs light at a particular wavelength, then
intensity of the sample beam will be less than that of the
reference beam. The motor light attenuator drives an optical edge
into the reference beam until the detector receives light of equal
intensity from sample and reference beam. The detector converts
IR radiation into electrical signals and sends it to recorder. The
output graph obtained is a plot of the wavelength of the entire
region versus the absorbance of the light at each wavelength.
A = log(I0/I)
The graph obtained is called as absorption spectrum
Components :
The modern IR spectrometers consist of light source,

sample chamber, Motor light attenuator, filter, detector, amplifier
and the recording devices.
Source of light :
 Near IR - Tungsten filament
 Mid IR - Nernst filament
– made of rare earth oxides
- Globar filament
– made of carborundum
 Far IR - High pressure mercury arc lamp
Function
Provides the wavelength region corresponding to IR
radiation.
Optical systems
 It is made of mineral salts (NaCl and KBr) transparent to
IR radiations
Function
The beam is guided and focused by mirrors silvered on
their surfaces.
Sample chamber (sample cell and reference cell)

 For Liquid sample
- Rock salt cell (NaCl cell, KBr cell )
 For Gas sample
- Cell with larger path length (NaCl or KBr cell)
 For Solid sample -
Pellet technique - KBr + Sample, made into paste
Mull technique - Nujol + Sample, made into paste.
Function
Used to hold the sample and reference materials.
Chopper
 Circular disc with alternate quadrants removed.
Function
Rotates and transmits sample and reference beam
alternatively to grating
Filters :
Monochromators - It consists of an entrance slit, a dispersing
element (prism or grating) and an exit slit.
 IR region – NaCl prism
 Far IR region – KBr prism
 Grating made of quartz or fused silica.
Function
Sends individual required frequencies to detector
Detectors :
 Thermocouple (EMF)
 Bolometer (ohms)
 Golay pneumatic cell
 Photo conductivity cell
Function
The detector converts the radiation signals into
current. The current is directly proportional to the concentration
of the solution.
Motor light attenuator

Function
Drives an optical edge into the reference beam until the
detector receives light of equal intensity from sample and
reference beam.
Amplifier and Recording system

 Pen/ digital recorder
Function
The output graph which is a plot of the wavelength of the
entire region versus the absorbance of the light at each
wavelength is obtained.
6.7 ATOMIC SPECTROSCOPY
Photometer
It is an instrument that furnishes the ratio or some
function of the ratio, of the radiant power of two electromagnetic
beams.
Spectrometer
An instrument with an entrance slit, a dispersing device
and one more exit slit with which the measurements are made at
selected wavelengths within the spectral range or by scanning
over the range. The quantity detected is a function of the radiant
power.
Spectrophotometer
It is a spectrometer with associated equipment, so that it
furnishes the ratio, or a function of the ratio, of the radiant power
of the two beams as a function of spectral wavelength. These two
beams are separated in time, space or both.
This technique involves the study of the absorption and

radiation by neutral atoms in the gaseous state. Thus in the AAS
the sample is first converted into an atomic vapor and then the
absorption of the atomic vapor is measured at a selected
wavelength which is characteristic of each individual element.
The technique is also called as absorption flame photometry,
because all the analyte applications of atomic absorption involve
spraying the solution of the sample into a flame. When a solution
having a mixture of metallic species is introduced into the flame,
the solvent evaporates and vapours of metallic species are
obtained. Some of the metal atoms can be raised to energy level
sufficiently high to emit characteristic radiation of metal – a
phenomenon that is used in flame emission spectrometry. But in
AAS the unexcited atoms serve as the basis for atomic absorption
measurements.
6.8 FLAME PHOTOMETRY
Principle
When a solution having a mixture of metallic species is
introduced into the flame, the solvent evaporates and vapours of
metallic species are obtained followed by the atomization. Some
of the metal atoms are excited sufficiently high to emit
characteristic radiation of metal i.e – a phenomenon used in
flame emission spectrometry.
COMPONENTS
NEBULISATION
 Pneumatic nebulisers.
Function
Filaments of liquid are drawn out from the bulk of the
solution. Liquid sample is drawn through the capillary by the
pressure differential generated by the high velocity gas stream
which passes over the sample orifice. This liquid is set into
oscillation. The filaments collapse to form droplets. The cloud of
droplets strikes an obstruction in the spray chamber termed a
splinter or impact bead. This breaks the larger droplets into
smaller ones. The aerosol (a very fine fog) is mixed with the
oxidant - fuel mixture and carried into the burner. Droplets larger
than about 20 microm are trapped in the spray chamber and flow
to waste.
FLAME
BURNER
 Turbulent flow or total consumption burner,
 Laminar Premix burner
 Non-flame atomizer.
Turbulent flow or total consumption burner:
Here the nebuliser and the burner are combined into a
single unit. The sample is drawn up the capillary and the fuel and
oxidant are carried through separate passages to meet and mix

with the analyte at the base of a flame.
Laminar Premix burner

The sample is aspirated by the stream of the oxidant. The
resulting aerosol is then mixed with fuel and then sent into the
flame.
Non-flame atomizer:
Instead of flame, electrically heating devices are used.
Function
As soon as the aerosol produced by nebulisation in the spray
chamber is transported into the flame the following sequence of
events occurs in rapid succession.
1. The solvent is evaporated, leaving minute particles of

dry salt.
2. The dry solids are converted into the gaseous state.
3. A part of all of the gaseous molecules is progressively
dissociated to give neutral atoms or radicals - the
atomisation step. These neutral atoms are the species
which absorb in AAS and AFS and are the potentially
emitting species on FES. The efficiency with which
the flame produces neutral atoms of the analyte is of
equal importance in each of the flame technique.
4. A portion of the neutral atoms may be thermally
excited by collisions with partially burnt components
in the flame gases, or even ionised. The fraction
excited is important in FES but a nuisance in AAS.
5. Some of the neutral atoms may combine with the
radicals in the flame gases to form new gaseous
compounds such as metal oxides. This gives rise to
the chemical interferences which are of nuisance in all
the three flame techniques.
FUEL GAS
 Natural gas, Propane, Butane, Acetylene or Hydrogen
could be used.
OXIDANT
 Air, Oxygen or Nitrous oxide could be used.
Fuel oxidant mix :

 For elements easily converted into atomic state, example
– Cu, Cd etc., low temperature flames are required which
is provided by Natural gas – air mixture.
 Elements forming oxides requires hot flame which are
provided by Acetylene - air mix especially for alkaline
earth metals and Nitric oxide – acetylene mix especially
for elements forming stable oxides like Al etc.
CHOPPER
 Circular disc with alternate quadrants removed
Function
The intensity of the source is made to fluctuate at a
constant frequency. This process is called modulation. Thus
rotation of the disc at constant speed provides a beam that is
chopped to the desired frequency, thus modulating the radiation
from the source.
FILTER
 Gratings are used.
Function
Minimize the flame background emission and resolve the
atomic emission lines from nearby lines and molecular fine
structure. Thus provides a narrow band width and isolate lines
that interfere with analysis.
SLITS
 Adjustable slits
Entrance slit is placed between the flame and monochromator

which permits only the radiation coming from the flame &
mirror.
Exit slit is kept between the monochromator and detector which
prevents the entry of interfering lines.
Function
To give greater intensity
DETECTOR and its Function

 The Photomultiplier tubes, Photo cell etc..
Function
It converts the radiation signals into electrical signals.
AMPLIFIER & RECORDER and their functions

Since the current coming out from the detector is weak,
amplifier is used to amplify the current and which is recorded
using a pen / digital recorder.
6.9 ATOMIC ABSORPTION SPECTROSCOPY
INTRODUCTION
This technique involves the study of the absorption and
radiation by neutral atoms in the gaseous state. Thus in the AAS,
when a solution having a mixture of metallic species is
metallic species are obtained and then the absorption of radiation
by the atomic vapor is measured at a selected wavelength which
is characteristic of each individual element. The technique is also
called as absorption flame photometry, because all the analyte
applications of atomic absorption involve spraying the solution of
the sample into a flame. In AAS the unexcited atoms serve as
the basis for atomic absorption measurements.
Principle
When a solution having a mixture of metallic species is
metallic species are obtained, followed by the atomization of the
vapor and then the absorption of radiation by the unexcited
atoms in the atomic vapor is measured at a selected wavelength
which is characteristic of each individual element. The flame
gases are treated as a medium containing free, unexcited atoms
capable of absorbing radiation from an external source. This
occurs when the radiation corresponds exactly to the energy
required for a transition of test element from ground electron
state to the excited state. The unabsorbed radiation passes
through a monochromator that isolates the exciting spectral lines
and into a photodetector. Absorption is measured by the
difference in transition signal in absence and presence of test
element.
 I 
log     cx
 I0 
 = absorption coefficient
x = path length
I = transmitted intensity
I0 = Incident intensity
c = concentration
RADIATION SOURCE
 Hollow cathode lamp with tungsten anode and cylindrical
analyte metal cathode in a glass tube with filler gas as
Ne or Ar, is used
Function
When a potential is applied across the electrodes
ionisation of the gas occurs and the current flows as the ions
migrate to the electrodes. If potential is large the gaseous cation
acquire enough kinetic energy to dislodge some of the metal
atoms from the cathode surface and produce an atomic cloud
called sputtering. A portion of the sputtered metal atoms are in

the excited state and thus emit their characteristic radiation.
Eventually the metal atoms diffuse back to the cathode surface
and redeposited. To remove interferences modulation is done.
CHOPPER
 Circular disc with alternate quadrants removed
Function
The intensity of the source is made to fluctuate at a
constant frequency. This process is called modulation. Thus
rotation of the disc at constant speed provides abeam that is
chopped to the desired frequency, thus modulating the radiation
from the source.
FUEL GAS
 Natural gas, Propane, Butane, Acetylene or Hydrogen
could be used.
OXIDANT
 Air, Oxygen or Nitrous oxide could be used.
Fuel oxidant mix :

 For elements easily converted into atomic state, example
– Cu, Cd etc., low temperature flames are required which
is provided by Natural gas – air mixture.
 Elements forming oxides requires hot flame which are
provided by Acetylene - air mix especially for alkaline
earth metals and Nitric oxide – acetylene mix especially
for elements forming stable oxides like Al etc.
BURNER
 Turbulent flow or total consumption burner,
 Laminar Premix burner
 Non-flame atomizer is used.
Turbulent flow or total consumption burner:
Here the nebuliser and the burner are combined into a
single unit. The sample is drawn up the capillary and the fuel and
oxidant are carried through separate passages to meet and mix

with the analyte at the base of a flame.
Laminar Premix burner
The sample is aspirated by the stream of the oxidant. The
resulting aerosol is then mixed with fuel and then sent into the
flame.
Non-flame atomizer:
Instead of flame, electrically heating devices are used.
Function
Vapourisation of the liquid sample occurs followed by the
atomization of the elements. The electrons of these elements
absorb energy from the light source and get excited from ground
state. This then emits the energy and returns to the ground state.
The unabsorbed radiation passes through a monochromator that
isolates the exciting spectral lines and into a photodetector.
FILTER
 Glass filters can generally be used
 Filters similar to the UV-Visible Monochromators could
be used.
Function
Provides a narrow band width and isolate lines that
interfere with analysis.
DETECTOR and its Function

The detectors that are used are same as in UV-visible
spectrophotometers, example the Photomultiplier tubes. Both null
point and direct reading meters calibrated in terms of Absorbance
or transmittance are used which receives alternating signals from
source and continuous one from the flame which are converted to
electrical signals. AC part of the signal is amplified and
unmodulated dc signal is ignored
AMPLIFIER AND RECORDER
Since the current coming out from the detector is weak,
amplifier is used to amplify the current and which is recorded
using a pen / digital recorder.

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Analytical Techniques 6.

To Recapitulate the Wave parameters

The unit for frequency is cycles per second or in (Hz).

Wave number is expressed as the waves per cm (cm-1). This unit

6.2 ELECTROMAGNETIC RADIATION

Electromagnetic radiation is a type of energy that is

The electromagnetic radiation is found to have dual nature

The principles and laws that govern the absorption of

Table 6.1 - The regions of electromagnetic spectrum

Type of transition resulting Frequency Type of Wavelength

When the monochromatic radiation passes through a

On integrating the equation (1), between the limits I = I0 at x = 0

where, k = absorption coefficient

The above equation is known as Lambert’s law.

Beer’s law (or) Beer-Lambert’s ’s law

When the monochromatic radiation passes through a

Let I0 be the radiant power of a beam incident upon a section of

The absorbance increases directly with the concentration

Thus Beer-Lambert’s law states that, “when a beam of

where , k = molar absorption coefficient.

On integrating the equation (3) between limits I = I0 at x = 0 and

(or) A  Cx ……. (4)

Equation (4) is called Beer-Lambert’s law.

Thus, the absorbance (A) is directly proportional to molar

Application of Beer- lamberts law

Determination of Unknown concentration of a solution

Then according to Beer-Lambert’s law

Let Cu be the unknown concentration of the solution which is to

From equation (1) and (2), we get

Limitations of Beer-Lambert’s law :

This law can be used only for

1) The transmittance of a solution is 30%. Determine the

Example 2: The solution of concentration of 2.5 10-4M has the

ii) The molar absorption coefficient

Example 3: The absorbance of a solution 1.2  10-4 M. is found

Example 4: A solution shows a transmittance of 20% when taken

Example 5 : A solution of thickness 2 cm transmits 40%

A monochromatic radiation is incident on a solution of 0.06 M

According to Beer-Lambert law,

6.3 INTRODUCTION TO SPECTROSCOPY

All chemical species interact with the electromagnetic

It is convenient to characterize absorptiometric methods

A white source (electromagnetic radiation) is focused on

A given sample continues to absorb energy and show an

A possible mechanism for this is by thermal collision. An

Another mechanism is that the energy gained from

The spectrum obtained could be either due to the excess

The study of spectroscopy can be carried out under the

Differences between Atomic and Molecular spectra

Higher energy level

Preparation of the coloured solution :

Example : The complexing agent ammonium hydroxide is added

Instrumentation for colorimetry

The amount of light transmitted depends on the depth of colour

Entrance slits: It provide a narrow source of the light.

Exit slit: It selects a narrow band of dispersed spectrum and

COLORIMETRY - Instrumentation Block diagram

LIGHT FILTER SAMPLE

MATERIALS USED IN COLORIMETRY

LIGHT SOURCE FILTER SAMPLE CELL

FUNCTIONS OF THE PARTS USED IN COLORIMETRY

Estimation of Concentration of a solution

Estimation of copper sulphate