UV-Vis Spectroscopy

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Outline

1. What is Spectroscopy?
2. UV-Vis Spectroscopy
3. Case study

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1
What is Spectroscopy?
The science which studies the properties
of matter, through its interaction with
electromagnetic radiation

The effect of interaction of EM radiation

with matter depends on the energy of
EM radiation.
Spectroscopy can be used for:
• Molecular identities
• Molecular conformations, geometries,
and sizes
• Chemical equilibria
• Rates of chemical reactions
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History
• Newton is traditionally regarded as the
father of spectroscopy
• In his book Optick, Newton demonstrated that
light could be split into its component colors by
means of prism, and these components could
be recombined to generate white light
• Kirchhoff & Bunsen recognized that each
atom and molecule has its own
characteristic spectrum
• This observation established spectroscopy as
a scientific tool for analyzing
the composition of materials.
• Later on scientist also analyzed light
coming from distant objects . 4
Electromagnetic radiation
Electromagnetic radiation is an electric and
magnetic disturbance traveling through space &
time
It has both particle & wave characteristics

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Types of Spectroscopy

Absorption Emission Scattering

Spectroscopy Spectroscopy Spectroscopy

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 Beer's Law

"The intensity of incident

monochromatic light decreases
with the concentration of
absorbing substance"
Lambert's Law

"The intensity of monochromatic

radiation as it passes through
transparent medium, decreases
with the thickness of the
medium"
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The change in intensity of
radiation due to absorption
can therefore be equated
as:

Absorbance is defined as the

amount of light which is
absorbed by the substance and is
calculated as the negative
logarithm of transmittance:

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•

9
•

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UV-Vis Spectroscopy

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 Introduction
• Ultraviolet-visible spectroscopy (UV-Vis ) refers to
absorption spectroscopy in the ultraviolet-visible spectral
region.
• In UV – vis spectroscopy, the EM rays passed to the
sample are absorbed by the , which increases the energy
of the system.
• This causes the excitation of an electron from a lower
energy state to a higher energy state.
• This excitation forms an absorption spectrum that can be
detected by the detectors in the spectrometer.
• The amount of photon (radiation) absorbed results in an
absorption spectrum which can then be measured in
terms of absorbance.
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• UV spectrum : Absorbance vs
wavelength, with peak around 217
nm.
• UV Range 100 – 400 nm
• Near UV : 250 – 400 nm
• Far UV : 190 – 250 nm
• Vacuum UV : <190 nm
• Vis Range 400 – 700 nm

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Energy levels

LUMO

HOMO

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Energy transitions
σ-σ*
• σ electron from orbital is excited to corresponding anti-bonding orbital σ*
• High energy transition, the entire region should be evacuated (Vacuum UV
region)
π- π*
• π electron in a bonding orbital is excited to corresponding anti-bonding orbital π*
• Compounds containing multiple bonds undergo π-π* transition.
n- σ*
• Saturated compounds containing one hetero atom with unshared pair of
electrons(n) undergoes n-σ* transition.
• This type of transition is very sensitive to hydrogen bonding
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n-π*
• An electron from non-bonding orbital is promoted to anti-bonding π*
orbital.
• This transition require minimum energy out of all transitions

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 UV spectrophotometer
Components of spectrophotometer
• Source
• Monochromator
• Sample containers
• Detector Output

• Computer
Reference Detector

Computer
Source

Monochromator Sample Detector

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• Source : Deuterium & Hydrogen lamps
• Monochromator/Wavelength selector
• Slits
• Mirrors
• Reflection grating
• Sample container (Cuvettes) : made of
quartz or glass
• Detectors : Two types
• Photomultiplier tube (PMT)
• Photodiode detector

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Detectors
Photomultiplier tube (PMT)
• It consist of photoemissive
cathode (emits electrons
when struck by photon),
anode (collects electron)
• These are very sensitive
• But intensity of light may
damage the PMT
Photodiode
• It consist of array of
multichannel photon detector
• These are less sensitive .
• They can measure fast
moving samples
19 15
 Single beam spectrophotometer
• All light passes through sample.
• Sample must be removed to measure the intensity of incident
radiation.
Output
Advantages
• Less expensive
• High sensitivity
Disadvantages
• Lack of compensation for circuit and voltage fluctuations. Computer
Source

Monochromator Sample/Cell Detector

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 Double beam spectrophotometer
• Chopper splits the light source into two beam
• Reference beam : For monitoring lamp energy
• Sample beam : Record absorbance from sample
Advantages Disadvantages
No circuit fluctuation Expensive and complex Output

Reference light

Computer
Source

Monochromator Sample/Cell Detector

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 Split beam spectrophotometer
• Beam splitter is used instead of Chopper
• Absorbance of sample and reference can be measured same time

Output

Reference Detector

Computer
Source

Monochromator Sample Detector

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Beam splitter
Advantages/Disadvantages & Applications of
UV Visible spectroscopy
Applications
1. Qualitative and quantitative analysis
2. Impurity detection
3. Unknown compound detection
4. Molecular weight calculation

Advantages Disadvantages
• Simple, inexpensive technique • Mixture analysis can be
• Can be used in qualitative and difficult.
quantitative for analysis • Not highly specific spectrum
for specific molecules 23
 Experimental Procedure to perform ULTRA-VIOLET VISIBLE
Contents SPECTROSCOPY
1. Aim
2. Apparatus &Chemical Used
3. Procedures
4. Solution Preparation
5. Experimental calculation
6. Factors that cause error

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Aim : To apply the Beer-Lambert relationship to an aqueous solution containing
an absorbing substance and thus determine its respective concentrations.

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Apparatus &Chemical Used
Apparatus:
1.Spectrophotometer 2. Cuvettes 3.Measuring flask (10mls.)
4.1mls and 10mls capacity pipettes 5. Pipette filler 6. storage bottle
7. Digital mass balance 8. Test tube rack 9. Beaker.
10. Stopper 11. Stop watch

Chemical/ Reagents:
1)Salicylic acid (0.1%)
2)Acetate buffer (0.05M)
3)Distilled water

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 Procedures:
1) We turn on the spectrophotometer and allow it to warm up for at least 20 min. Then we
determine the absorption spectrum using the standard acetate buffer.
2) We select one of the cuvettes for the blank solution (in this case acetate buffer) and we do
not interchange it with the other cuvettes also we did not handle the lower portion of
cuvettes through which the light passes.
3) We always rinse the cuvettes with several portions of solution by using acetate buffer
before taking a measurement.
4) Then we wipe the outside of cuvettes with tissue paper.

1) Then we inserted the cuvette into the cell holder with the index line facing us to avoiding
scratching.
6)Then we turn the wavelength control knob to 265nm with blank solution calibrate the
spectrophotometer.

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Solution preparation
A. Preparation of the solution
1) We prepared a stock solution of 0.1% salicylic acid by: accurately measuring 10mg of
salicylic acid by digital mass balance then we dissolved the obtained mass in 100ml of
acetate buffer in beaker.
2) From that stock solution above we prepared serial dilution containing 0.05%, 0.025%,
0.01%, 0.005%, 0.0025, and 0.001%.(Note all percentages are in weight by volume ).

• We apply dilution law of same substance as:

• C1 V1=C2V2
• Where:
•C1=concentration of concentrated (stock) solution. V1= volume of

concentrated (stock) solution.

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C2= concentration of diluted solution. V2= volume of diluted solution.
Note: V2 will be equal to 10ml because we have to diluting the stock solution by adding sufficient amount of
acetate buffer to make a solution of 10ml in measuring flask. Also C1 and C2 known then V1 obtained by
following formula V1 = C2V2/C1
3) Then we labeled test tube from 1 up to 6. After obtained required volume of stock solution to be taken
from beaker containing stock solution of salicylic acid. Then we add sufficient amount of acetate buffer
to make 10ml solution results summarized in following table.
4) Then we prepared (standard blank) in which 20ml of acetate buffer is substituted for the sample.

TABLE.1;
S/N 1 2 3 4 5 6
Concentration 0.05 0.025 0.01 0.005 0.0025 0.001
(%w/v)
Volume of stock 5.00 2.50 1.00 0.50 0.25 0.10
solution need(ml)

Volume of diluted 10 10 10 10 10 10
solution(ml)

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 B. Determination of absorbance Calculation
1) We set blank solution in the cuvette and calibrate at wavelength at wavelength control of 296 nm.
2) Then we inserted cuvette containing the sample- then we read and recorded the absorbance for all solutions in
above section A.
3) Then we determine the absorbance of unknown sample.

Concentration in mol/dm3:
Since
Yg in 1000 cm3

Xg in 100 cm3

Since X is mass of sample in above table then mass Y which is in 1000cm3 can be obtain.
But molar mass of salicylic acid= 138g/mol. 33
Then we change those concentrations in w/v% in Table 1 above into Molarity Then we obtain Table below.
TABLE .2;

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 5) From the graph we obtain the equation below:

We plot the graph of Absorbance vs. concentration in Excel sheet as shown below:
Y = 3.7129X + 0.006
Hence the unknown concentration can be obtained
ABSORBANCE VS. CONCENTRATION (MOLE/LITRE) since:
0.025 Y=value from y-axis i.e. the absorbance
y = 3.7129x + 0.006 X=value from x-axis i.e. the concentration in Mol/litre
0.02
R² = 0.8294
But absorbance (Y value) of unknown sample is
obtain which is equal to 0.042 Then we can find the
0.015
Absorbance

value of unknown concentration!

0.01
Take
0.005 0.042=3.7129X+ 0.006
Then
0 0.042-0.006=3.7129X
0 0.0005 0.001 0.0015 0.002 0.0025 0.003 0.0035 0.004
concentration (mol/litre) 0.036=3.7129X
Then divide by 3.7129 both side to obtain X=9.696x
10-3 There fore
Unknown concentration in mol/litre of solution is
equal to 9.696x 10-3

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 6) The molar extinction coefficient of SA
From below equation

A = ϵ𝑐 𝑏
Where:
A= absorbance
ϵ=extinction coefficient (mol-1dm3cm-1) b= path length of cell (cm)

c= concentration of absorbing species.

Take
A1 =0.007 and A2=0.003
Also
C = 3.62x 10-4and C =1.81x 10-4
1 2
b =1cm
A1=ϵ1c1b and A2=ϵ2C2b
0.007= ϵx
1
3.62x 10-4x1cm …………………………………….eqn.1
0.003= ϵ2x1.81x 10 x1cm…………………………………………eqn.2
-4
Then

ϵ1=19.33 ϵ2=16.57

Then molar absorbivity of SA Will be 19.33+16.57=35.9 35.9/2=17.95moll-1cm-1

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 Factors that cause Errors
Source of errors:
• Stray Light: A problem when working at limits of a spectrometer’s range.

• Cells and Solvents: Everything else except the sample should be as

transparent as possible.

• Sample Preparation: If two samples are prepared so that one carries along
greater concentration of insoluble particulates, then additional scattering will
lead to an apparent greater absorption.

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4) Slit Width Affects Absorbance Measurements: If a significant variation in
absorptivity occurs over the spectral bandwidth admitted by the slit, a non- linear
variation (non-Beer’s Law) with concentration will be observed. This arises
because the spectrometer measures the average transmissivity over the spectral
bandwidth, but transmissivity and concentration are not linearly related. Keep slit
width large to increase S/N ratio, but must keep it small enough to maintain a linear
relationship with concentration changes. This effect is minimized if the absorptivity
changes slowly with wavelength. Select a wavelength near a peak maximum. Use a
slit width to provide a bandwidth about 1/10 the spectral feature width.

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5) Wavelength error: In liquids, the extinction coefficient usually changes slowly with
wavelength. A peak of the absorbance curve (a wavelength where the absorbance
reaches a maximum) is where the rate of change in absorbance with wavelength is
smallest. Measurements are usually made at a peak to minimize errors produced by
errors in wavelength in the instrument, that is errors due to having a different
extinction coefficient than assumed.

The Beer-Lambert Law will not be obeyed if

• The photons of light striking the detector do not all have an equal chance of absorption by the sample. This can
happen if they have different absorption coefficients, different path lengths through the sample, or if they
encounter different concentrations of sample molecules. Also

• If anything else is present in the sample that absorbs light or causes light scattering, the measured absorbance will
not be zero when the analyte's concentration is zero, contrary to Beer's Law.

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• If the absorber undergoes any type of chemical reaction or equilibrium that varies as a function of
concentration, Beer's Law will not be obeyed with respect to the overall or total concentration,
because the concentration of the actual absorbing molecule is not proportional to the overall
concentration of the solution. The "c" in Beer's Law refers to the concentration of just the
absorber, not to the total concentration of all the compounds reacting with or in equilibrium with
the absorber. Even if Beer's Law holds exactly for each individual compound, the total
absorbance of the mixture will not follow Beer's Law with respect to the total concentration if the
proportion of each compound changes with concentration (unless by chance the absorptivity of
all those compounds happens to be exactly the same).

• Deviations in absorptivity coefficients at high concentrations (> 0.01M) due to

electrostatic interactions. Changes in refractive index at high analyte concentration.

• Fluorescence or phosphorescence of the sample and Non- monochromatic radiation.

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 A Case study : Suspension stability of Titania NanoParticles
(Ti02) Studied by UV-VIS spectroscopy Method.
Aim : To explore the dispersion and stability of titania nanoparticles in an aqueous media with
different types of dispersants by using the Ultraviolet–Visible (UV–Vis) spectroscopy
• Hydrochloric and nitric acids as well as pure ammonia were used to determine the stability of the
suspension in the acidic region (pH=2.5) and basic area (pH=9.5), respectively.
• In addition, for measuring sustainability of suspension and creating steric, and electrosteric
repulsive forces, ethylene glycol and ethylene glycol plus ammonia were employed, respectively.
• UV–V is spectrometry was applied to realize the effect of nano titania concentrations and different
types of dispersants of samples containing different amounts of nano titania and different types of
dispersants on stability of TiO2-containing suspensions.

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• Nano sized titania is produced worldwide in large quantities because of its unique properties for
applications such as pigments, cosmetics, paper, fine ceramics, as well as photocatalysts for
environmental purification, catalyst supports, and dielectric materials . Because Owing to the very
strong Van-der-Waals interactions, TiO2 nanoparticles almost incline to form aggregates or hard
agglomerate.

• For the stable dispersion of TiO2 nanoparticles in water is such a significant challenge.

• To evaluate the stability of ceramic suspensions is zeta potential measurement which is widely used
as an important criterion . A high absolute value of zeta potential is attributed to a well-dispersed
suspension. According to the zeta potential values, the isoelectric point (IEP) of titania nano powder
is about 4.5 - 6.0. However, to access a stable suspension of ceramic particles, the slurry needs a
zeta potential variation of about ± 40 eV from IEP. Therefore, the optimized conditions to produce a
stable suspension of nano titania with electrostatic mechanism is either in pH range of 2.5-3.5 and
9.5-10.5; i.e. in acidic pHs and basic ones, respectively.

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Table 1. Stability time of TiO2 suspensions with variable concentrations of TiO2

Stability time (min)

HCl HNO3 NH3

Without NH3+EG
TiO2 Additive (pH=2.5) (pH=2.5) 5%wt EG (pH=9.5)
(pH=6.5) (pH=9.5,
Concentration
5% EG)

0.25
27 91 112 240 360 720

0.5
24 76 91 192 315 660

0.75
21 65 73 141 282 630

1 18 56 64 105 210 612

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• The proper amounts of dispersant was added to 100 ml distilled water followed by the addition
of 0.25, 0.5, 0.75 and 1g of nano titania. pH of suspension was adjusted to a desired value, then;
the suspension was stirred for 30 min using a magnetic stirrer.
• Fig. 1 depicts sediment time of nano titania suspensions with different dispersants as a function of
titania concentration. Comparison of Fig. 1 (a) and (b) showed that increasing titania
concentration gave rise to the reduction of stability time in all samples.
• The reason lies in this fact that OH- groups in basic media have larger space size than the H+
groups in the acidic region. Therefore, in spite of existence of repulsive forces which cause an
electrostatic stability, the above fact generates an extra steric repulsion which accompanies higher
stability times in the basic media.
• As shown in Fig. 1(b), the sample prepared by NH3+EG displayed the highest stability time
compared to other suspensions. In other words, electrostreic repulsive mechanism in the sample
stabilized with NH3+EG has the highest efficiency to stabilize the nano titania suspension in
comparison with steric and electrostatic mechanisms.

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Fig. 1. Stability time versus TiO2 concentration with different type of additives
(a) electrostatic mechanism (b) steric and electrosteric mechanisms
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Fig. 2. Appearance of TiO2 suspensions after one day.
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Fig. 3. UV-Vis
absorption spectra
of different
stabilized (a)
electrostatic (b)
steric and
electrosteric samples
after soaking time of
14 days

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• Fig. 3 As can be seen in the figure, the absorption is dependent on not only the wavelength but also the type of dispersant.
The diversities of UV–Vis absorption between the samples containing additives and one with no additive demonstrate the
dispersion efficiency in different samples.

• In other words, the additives have caused the nano titania particles to segregate from each other and has prevented them
from forming agglomerates, so that a higher surface area of TiO2 particles after dispersion could be achieved. It seems that
the combination of NH3 and EG has led to the better consequences. Through this, the total area of titania particles which
exposed to UV radiation were increased, resulting in more excitations. Hence, by improving the quality of the dispersion,
a higher absorption in the UV–Vis spectra is expected.

• The highest absorption of ultraviolet radiations in suspension stabilized by NH3+EG, confirmed the higher efficiency of
electrosteric repulsive forces in comparison with electrostatic and steric mechanisms. This behavior is attributed to this
fact that the absorbed ions in electrostatic mechanism form a charged layer around the particles preventing the
aggregation, while in steric mechanism, on the other hand, a volume restriction component as well as a mixing or osmotic
component are the two influential factors on stabilization of suspension . Therefore, the electrosteric stabilization, due to
possessing both advantages of electrostatic and steric stabilizations, exhibits a more efficient role in stabilization of
suspensions compared to the other stabilization mechanisms.

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Fig. 4. UV–Vis
absorption of
samples with
electrostatic steric
and electrosteric
stabilized
mechanisms in
comparison with no
stabilized sample in
= 300 nm.

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Infrared Spectroscopy

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IR spectroscopy

• IR radiation does not have sufficient energy to induce

electronic transitions.
• If the energy of IR region concedes with the natural frequency
of vibration of molecule then it undergoes into vibrational
transitions.
• For a molecule to absorb IR radiation, it must be accompanied
change in dipole moment.
• IR spectroscopy gives information about functional groups in a
compound.
• Every bond or portion of a molecule or functional group
has characteristic frequency of absorption.
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• The vibration of two atoms connected with covalent bond
behaves like two vibrating ball connected by spring.

• Infrared spectroscopy also known as Vibrational spectroscopy

is the study of absorption of IR radiation, which leads
to vibrational transition.
• The change in vibrational level depends on
• Mass of atom present in molecule
• Strength of bonds
• Arrangement on atoms within molecule
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Effect of absorption
Molecule absorb incident radiation and its energy increases, there are
four possibilities
1. Rotational Transition
2. Vibrational transition
3. Electronic transition
4. Translational transition

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 Symmetric
Stretching
Vibrations
Asymmetric

Types of Scissoring
Vibrations In Plane
bending
Rocking
Bending
Vibrations
Wagging
Out of Plane
bending
Twisting 54
IR spectrum
•IR spectrum is plotted between
Transmission and wavenumber, each
dip is called band or peak.
•IR spectrum has peaks and bands
• Peaks
• Weak
• Medium
• Strong
• Bands
• Broad
• Sharp 55
 Fingerprint region
• The fingerprint region is the frequency range
between about 1450 and 600 wavenumbers
that contains a large number of stretching
and bending vibrations.
• Here bending vibrations are usually more
than stretching vibrations
• Many compounds shows unique absorption
bands in this region.

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Factors influencing Vibrational frequency
• Symmetry : Since symmetric molecules have zero dipole moment, so
they are IR inactive.
• Fermi resonance : Results in unexpected shift in energy of the bands
• Hydrogen bonding : Result in downwards frequency shift.
• Electronic effects : effects such as Induced, mesomeric & field may
change in absorption frequency.
• Bond angle : Different bond angles also affect the absorption
frequency.

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 Instrumentation
• Similar to UV, IR spectrophotometer have similar instrumentation.
• IR source
• Monochromator/Wavelength selector
• Sample cells Output
• Detectors
• Computer
Reference Detector

Computer
Source

Monochromator Sample Detector

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IR Radiation source
• IR source must be steady, & produce intensity enough to detect
• Some common IR sources
Nernst Glower 1 – 50 µm Mid IR

Glober 1 – 50 µm Mid IR

Tanustan lamp 0.78 – 2.5 µm Near IR

Mercury arc Lamp 50 – 300 µm Far IR

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Monochromator/Wavelength selector
• It is made up of
• Slits
• Mirrors
• Reflection grating
• Grating is the arrangement of
ruling on some material. The rays
which are incident on grating gets
reinforced with reflected rays.

Sample cell
Material containing sample must be
transparent to IR radiation
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 Sample preparation
Sample may be liquid, gas or solid different phases have
different sampling techniques.

Liquid sampling Gas sampling

• Gas samples are introduced into
• Liquid sample is sandwiched in a glass cell made up of NaCl
between two alkali halide plates
• The sample cell thickness is
around 0.01 - 0.05 mm

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Solid sampling
1. Solid run in solution :
• Solid is dissolved in non
aqueous solvent
• A drop of solution is placed
over alkali metal disk, let it
evaporate
• The obtained film is then
mounted
on spectrophotometer
2. Mull Technique :
• Finely powdered sample added
with mulling agent (Nujol) to
obtain a thick paste (mull)
• Mull is then placed in between
two plates, thickness is
adjusted
• Plates are then analysed
on spectrophotometer
3. Pressed pellet technique :
• Finally powered sample + KBr
• Mixed in a vibrating ball mill
• The mixture is then compressed
at very high pressure to form
small pellet.
• The pallet is then analysed
in spectrophotometer.
4. Solid film :
• Amorphous solid is dissolved in
volatile sample
• The solution is poured into NaCl
plate
• By heating volatile solvent is
evaporated
• Sample plated is then analysed 62
 Detectors
Thermocouple Dissimilar metal junction Cheap, slow, insensitive
(Thermoelectric effect)
Bolometer Ni, Pt resistance Highly sensitive
thermometer
Golay cell Metal cylinder with Xenon Faster, wide wavelength
gas range
Pyro electric Triglycine sulfate Fast and sensitive (mid IR)
piezoelectric material
Photoconductive PbS, CdS, light sensitive cells Fast and Sensitive (Far IR)

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Advantages/Disadvantages & Applications of
IR Visible spectroscopy
Applications
1. Identification of functional groups
2. Studying kinetics of reaction
3. Impurity detection

Advantages Disadvantages
• Simple, inexpensive technique • IR alone can’t determine
• Can be used in qualitative and structure of molecule
quantitative for analysis • Can’t detect non polar
molecules 64
 IR SPECTROMETER

DISPERSIVE FTIR

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 Dispersive IR spectrophotometer
● Older technology, used in 1st and 2nd
generation IR spectrophotometers.
● Incident IR radiation is dispersed by a
monochromator (prism,gratings and slits)
into its component frequencies.
● Monochromator has two fixed mirrors which
produces only constructive interference.
● The detector measures the amount of
energy at each frequency which has passed
through the sample.
● This results in a spectrum which is a plot of
intensity vs. frequency.

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Fourier Transform Infrared (FTIR) Spectroscopy
•FTIR is the extension of IR spectroscopy
•In IR, radiation is analyse as individual frequencies using
monochromator, While in FTIR all frequencies are are analyses
simultaneously.
•Instrument used for FTIR is Michelson interferometer
•The interference pattern obtained is analysed using mathematical
tool "Fourier transform".
•It obtains an infrared spectra by first collecting an
interferogram of a sample signal using an interferometer, then
performs a Fourier Transform on the interferogram to obtain
the spectrum of absorption or emission of solid, liquid or gas.
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How the FTIR works?
• IR 68
Spectroscopy is a qualitative analytical technique that helps to indicate
mainly the functional group of a molecule.
• When the IR radiation irradiated to the molecule the part of molecule which is
functional absorbs it, as a result of this absorbance the molecular vibration
increases.
• After the excitation of molecular vibration, these molecules comes back to
their original state by releasing that energy of certain wave number which is
recorded as transmittance on the spectrophotometer.
• IR is low energy, low frequency and long wavelength radiation with low wave
number that can only cause an increase in molecular vibrations (i.e. stretching
& bending).
• So by triggering molecular vibrations through irradiation with infrared light
provides mostly information about the presence or absence of certain
functional groups.
 FTIR spectrophotometer working

● Interferometers are used in place of

monochromators.
● A typical interferometer consists of
two perpendicular mirrors and a
beamsplitter. One of the mirror is a
stationary mirror and another one is a
movable mirror.
● We can analyse entire spectrum at a
time using interferometer.

69
 Interferometer working

● The beamsplitter is designed to transmit half of the light and reflect half of the
light.
● The reflected light strike the stationary mirror and transmitted light strikes movable
mirror, and gets reflected back.
● Two beams of light recombine with each other at the beamsplitter.
● If the distances travelled by two beams are the same which means the distances
of two mirrors from the beamsplitter are the same, the situation is defined as zero
path difference (ZPD).
● If the movable mirror moves back & forth from the beamsplitter, the light beam
which strikes the movable mirror will travel a shorter/longer distance than the light
beam which strikes the stationary mirror.
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● Let Δ be the distance travelled by movable mirror, then extra distance
travelled by light will be 2Δ.
● The extra distance is defined as the optical path difference (OPD) and is
represented by delta.

If OPD = n.λ

If OPD = (n+1/2).λ

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● If there is neither constructive nor destructive interference i.e.
n.λ < OPD < (n+½).λ
● Then the intensity of exit the signal will be in between maximum and minimum
● Since the mirror moves back and forth, the intensity of the signal increases
and decreases.
● So in FTIR instead of analysing single wavelength at a time, we can analyse
all the wavelengths simultaneously.
● Since the analysis required the Interpretation of spectrum of different
frequencies, so this interpretation is done with the help of mathematical tool
called Fourier transformation.

72
Dispersive IR vs FTIR

Properties Dispersive IR FTIR

Moving components Many moving components Only mirrors

Calibration Manual calibration against Internally calibrated

reference is required

Stray light Stray light may cause Stray light does not affect
spurious reading detector
Scan speed Slow Fast

Resolution Small amount of IR beam is Much larger beam aperture :

used : Moderate resolution Improved resolution
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Application
74

• Identification of inorganic compounds and organic

compounds.
• Identification of components of an unknown mixture
• Analysis of solids, liquids, and gasses
• Detection of impurities in compound
• In remote sensing
• In measurement and analysis of Atmospheric Spectra
- Solar irradiance at any point on earth
- Longwave/terrestrial radiation spectra
• Can also be used on satellites to probe the space
 Advantages of FTIR

● Speed: Because all of the frequencies are measured simultaneously, most

measurements by FT-IR are made in a matter of seconds rather than several
minutes.
● Mechanical Simplicity: The moving mirror in the interferometer is the only
continuously moving part in the instrument. Thus, there is very little possibility of
mechanical breakdown.
● Very reliable technique.
● No calibration required

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Disadvantages of FTIR

• The sampling chamber of an FTIR can present some limitations due to its
relatively small size.
• Mount pieces can obstruct the IR beam, Usually only small items as rings can
be tested.
• Several materials completely absorb infrared radiation, consequently, it may
be impossible to get a reliable result.

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 Case study 1: Application of
Fourier-Transform Infrared Spectroscopy (FTIR) for
the Study of Cultural Heritage Artifacts
• The FTIR spectroscopy can be a valuable tool for the analysis of
different types of historical artifacts due to its sensitivity,
specificity and non-destructive character
• Technique can be applied for characterization of metallic artifacts
(such as iron, bronze objects), paper artifacts, ancient ceramics &
historical textiles
• Chemical changes because of materials ageing can be observed
and sometimes quantified using FTIR spectroscopy
Aim of study
To identify different fibers which are used in given artifact.
77
 Sample
• The analyzed textile artifact represents a traditional
pillowcase originating from Moldavia historical region,
manufactured at the end of the 19th century –
beginning of the 20th century.
• From the large artifact, several smaller samples were
obtained, representing both the support material and
the colored fibers.

Analysis
• The FTIR spectra were recorded on a FTIR 6300
instrument.
• All the spectra recorded in the range 4000–400 cm−1
were averaged over 32 scans.

78
79
 Results

• As it can be observed from the spectrum ,the signals are specific for the
textile support without interferences due to the presence of different dyes
onto the surface of the fibers.
• At 1634 and 1511 cm−1 superimposed spectra of the colored fibers showed
strong bands corresponding to amide I and amide II.
• Together with several other intense characteristic bands (1390 cm−1 – CH3
symmetrical deformation and 1055 cm−1 – asymmetrical stretch C-O-C and
C-N ), confirming that the textile support is wool in all the cases. 80
• Spectra of cotton, flax and hemp recorded and compared with
artifact material.
• The FTIR spectrum of the white sample, which is the main material of the
artifact, reveals the presence of the characteristic peaks corresponding to
cellulose and lignin and apparently may be hemp or flax.
• The shape of the sample spectrum is closed to the flax spectrum
81
• The average values of the band intensity ratios R1 (1595/1105) and
R2 (I1595/I2900) were calculated for three types of pure cellulosic
fibers (cotton, hemp and flax) and for the support sample.

82
 Conclusion
• Despite minor differences in different types of cellulosic fibers, FTIR
technique proved to be a useful tool to distinguish them on the basis
of some of their structural features.
• Comparison between spectra of different cellulosic fibers,
the intensity ratios can lead to valuable results which allow
differentiation even in the case of problematic species such as flax
and hemp.
• The obtained results prove that artifacts is formed of two types of
fibers: the support textile fiber, identified as flax, and the colored
fibbers, identified as wool fibers dyed with natural colorants.

83
 Case study 2:Material susceptibility towards
Vikane
• Vikane (SO 2F 2 , Sulfuryl fluoride) used against drywood termites &
other domestic pests.
• It has several advantages
• Easy dispersal into structure
• Rapid penetration into material
• Negligible residue formation

Aim of case study :

Examine the extent of absorption of Vikane on different material.

84
Method used..
• Vapor-phase depletion measurements ,fundamental method to
determine the interaction of a solid material with a gas (Vikane).
• When a gas is placed in a closed chamber with a test material, the
only decrease in the gas concentration will occur due to
• Leakage or penetration through seals or gaps in the chamber
• Absorption or reaction with the chamber walls
• Absorption or reaction with the test substrate surface
• Thus by controlling different parameters, Different materials may be
compared for their specific absorption and reactivity to the gas.

85
 Experimental setup
• The concentration of Vikane was measured in situ
with IR spectroscopy.
• Substrate was placed in IR gas cell, Vikane was
introduced, & concentration of Vikene
was measured for certain amount of time .
• The test materials had an exterior dimension of 2.5
x 5.0 cm.
• 2-3 ml of 100% Vikane was injected into the gas
cell, and the automated IR collection program was
initiated.
• 8 hours later, spectra were taken to determine
whether the test materials were absorbing Vikane.
• The intensity of IR absorbance band is proportional
to concentration of species

86
Result
The IR absorption of Vikane at 1492 cm-1 versus collection time.

• Depletion curve at 0% RH.

• For the blank cell, the Vikane concentration
decreases slightly, then remains constant.
• Similar pattern was observed for metal surfaces
• Vikane concentration depletion for cotton was 5%.
• In case of silk 37% decrease in Vikane
concentration was observed
• Depletion curve at 65% RH.
•For metals Viken absorption increases .
•But in case of organic surfaces such as cotton, silk
absorbance of Vikane decreases.
87
Conclusion
• Depletion studies showed that IR spectroscopy is a useful tool for
monitoring changes in the gaseous environment in an IR cell.
• While it is not possible to determine from these experiments whether
the Vikane was reacting with the materials or not.
• For metallic surfaces relative humidity affects the absorption of
Vikene, while In case of organic samples relative humidity does not
affects the absorption.

88
RAMAN SPECTROSCOPY

89
 CONTENTS
 INTRODUCTION
 BASIC PRINCIPLES
 INSTRUMENTATION
 APPLICATIONS
 CASE STUDY

90
➢ Sir Chandrasekhara Venkata Raman
- November 7, 1888 - November 21, 1970
- Won the Nobel prize in 1930 for Physics and got Bharat Ratna Award
in1954
- Discovered the “Raman effect”
- Besides discovering the Raman effect he studied extensively in X-ray
Diffractions, Acoustics, Optics, Dielectrics and Colloidal solutions.

91
 INTRODUCTION

Spectroscopy:- Spectroscopy is that branch of science which deals with

the study of interaction of electromagnetic radiations with matter.

Raman spectroscopy is the measurement of the wavelength and intensity of

inelastically scattered light from molecules.
The Raman scattered light occurs at wavelengths that are shifted from the
incident light by the energies of molecular vibrations.
Raman spectroscopy is used to determine the molecular motions, especially
the vibrational one.

92
• Raman spectroscopy is among the handful of catalyst characterization techniques
that can provide molecular-level information.

A typical Raman System

93
 BASIC PRINCIPLE

Scattering of Light :-When light passes from one medium to any other medium say
air, glass or water then a part of the light is absorbed by particles of the medium
preceded by its subsequent radiation in a particular direction. This phenomenon is
termed as Scattering of light. The intensity of scattered light depends on the size of
the particles and wavelength of the light.

Scattering of Light

Elastic collision Inelastic Collision

94
Elastic collision Inelastic Collision
(No exchange of Energy) (Exchange of Energy)

Rayleigh Scattering Raman Scattering

Rayleigh scattering is the phenomena of scattering of The scattering of radiation with change in frequency
light particles majorly by the molecules of gas is called Raman Scattering.
(sometimes also by solid and liquid). This scattering of
light was first noticed by Lord Rayleigh in 1871
Inelastic: collision between photon and
molecule results in a net change in energy
Elastic: collision between photon and
molecule results in no change in energy

95
96
(Nobel Prize in Physics 1930)

97
 Energy Scheme for Photon Scattering

Virtual
State

h 0 h 0 +h m

h h h
Energy

0 0 0
h m

E0+h m

E0
IR Rayleigh Stokes Anti-Stokes
Absorption Scattering Scattering Scattering
(elastic)
Raman (inelastic)

The Raman effect comprises a very small fraction, about 1 in 107 of the incident photons.

98
mm

◼ Atoms are at a certain energy level at any

given time.
◼ As a laser light hits the atom, it is excited
and reaches a higher level of energy, and
then is brought back down.
◼ If an atom is at a given energy level, it can
be excited then fall below the original
level.
◼ Anti-stokes spectrum are mirror
spectrums of Stokes Raman Spectrums

99
Classical theory of raman effect

10 100
 Condition for raman spectroscopy

Raman spectra occurs as a result of oscillation of a dipole moment, induced in

a molecules by the oscillating electric field of an incident wave.

As the induced dipole moment is directly proportional to the polarisability of

the molecules, the molecules must possess anisotropic polarisability which
should change during molecular rotation or vibration for vibrational or
rotational-vibrational raman spectra.

Anisotropic polarisability depends upon the orientation of the molecules.

In the presence of an electric field, the electron cloud of an atom or
molecules is distorted or polarised.

10
1
What Exactly is being Measured?

When Light hits a sample, It is Excited, and is forced to

vibrate and move. It is these vibrations which we are
measuring.

102
 Raman Spectrum
A Raman spectrum is a plot of the intensity of Raman scattered
radiation as a function of its frequency difference from the
incident radiation (usually in units of wavenumbers, cm-1). This
difference is called the Raman shift.

103
 RAMAN INSTRUMENTATION
There are following component involves:-
1. Laser or source of light
2. Filter
3. Sample holder
4. detector

10
4
The block design dispersive Raman scattering system:

Wavelength Detector Recorder

Recorde
Recorde
Sample selector In Ga As or rr
GE

90·

Radiation
sources

Block diagram

10
5
10
6
 1. Laser or source of light

• Lasers are generally the only source strong enough to scatter lots
of light and lead to detectable raman scattering.
• Lasers operate using the principle of stimulated emission.
• Electronic population inversion is required to achieve gain via
stimulated emission (before the fluorescence lifetime is reached)
• Population inversion is achieved by “pumping” using lots of photons
in a variety of laser gain media

10
7
List of Various laser source
S.No. Laser wavelength
01 Nd:YAG 1064nm

02 He:Ne 633nm

03 Argon ion 488nm

04 GaAlAs diode 785nm

05 Co2 10600nm

06 Ti-Sapphire 800nm

10
8
A :- He:Ne laser
• Filled with 7:1 He & Ne gas optimum output of 6328 Å
• High voltage excitation is preferred
B :- Nd:YAG System
• A typical laser system –the neoAdymium-doped yttrium
aluminum garnet or Nd+3
• YAG is a cubic crystalline material
• Crystal field splitting causes electronic energy level splitting
• Nd: YAG laser are optically pumped using a flash tube or laser
diodes.
• These are the one of the most common type of laser.
• It emits 1064 nm wavelength
10
9
 2.Filter
• It is therefore essential to have monochromatic
radiations.
• For getting monochromatic radiations filters are used.
• They may be made of nickel oxide glass or quartz glass.
• Sometimes a suitable colored solution such as an
aqueous solution of ferricyanide or iodine in CCl2 may be
used as a monochromator.

11
0
 3.Sample holder
• For the study of raman effect the type of sample holder to be used
depends upon the intensity of sources ,the nature and availability
of the sample.
• The study of raman spectra of gases requires samples holders
which are generally bigger in size than those for liquids.
• Solids are dissolved before subjecting to raman spectrograph.
• Any solvents which is suitable for the ultraviolet spectra can be
used for the study of raman spectra.
• Water is regarded as good solvents for the study of inorganic
compounds in raman spectroscopy.
11
1
 4.Detector
• Researchers traditionally used single points detectors such as
photocounting, photomultiplier(PMT), not because of the
weakness of a typical raman signal, longer exposure times
were often required to obtains raman spectrum of a decent
quality.
• Now days multichannel detectors like photodiode arrays(PDA),
charged couple devices(CCD)
• Sensitivity & performance of modern CCD detectors are high.

11
2
 APPLICATION
Pharmaceuticals and Cosmetics:-
• Compound distribution in tablets
• Powder content and purity
• Raw material verification
Geology and Mineralogy
• Gemstone and mineral identification
• Mineral and phase distribution in rock sections
• Mineral behavior under extreme conditions
Carbon Materials
• Purity of carbon nanotubes (CNTs)
• Hard disk drives
• Defect/disorder analysis in carbon materials
11
3
Semiconductors
• Purity
• Alloy composition
• Superlattice structure
Life Science
• DNA/RNA Analysis
• Drug/cell interactions
• Bone structure

114
EXPERIMENTAL EQUIPMENT

115
 CONCLUSION
• The portion of the Raman spectra between wavenumbers of
465–830 cm-1, which represent the main features belonging to
the manganese oxides, were used for the CLS (Classical least
square fitting) analysis, shows the raw Raman spectra and their
corresponding cryptomelane scores returned by the CLS
analysis. A Linear combination of the scores of the cryptomelane
and amorphous manganese oxide and their corresponding
loadings spectra.

116
 CASE STUDY-2

Case:- Modifying alumina with CaO or MgO in supported Ni and Ni–Co catalysts and its effect on dry
reforming of CH4.

Objective:-Modifying the support of the alumina (Al2O3) supported Ni and Ni–Co catalysts with
limited amounts of CaO and MgO had an effect on the catalytic activity during the reforming of CH4
with CO2 at 873 K.

To achieve the above objectives modified-Al2O3 supports and modified-Al2O3 supported Ni and
Ni–Co catalysts were synthesized. The total metal loading (Ni or Ni + Co) and Ni:Co ratio, where
applicable, was kept constant. The synthesized catalysts were then characterized for their surface
area, and by ultraviolet– visible (UV–vis) spectroscopy, H2-temperature programmed reduction
(H2-TPR), X-ray diffraction (XRD), H2-temperature programmed desorption (H2-TPD), CO2-
temperature pro-grammed desorption (CO2-TPD), and temperature programmed hydrogenation
(TPH) studies. The Al2O3 and modified-Al2O3 supported Ni and Ni–Co catalysts were then tested
for the DRM reaction. The nature and amount of carbon deposited over the spent catalysts were
also studied by thermogravimetric analysis (TGA) and Raman spectroscopy. Based on these results
the changes in the catalytic performance are discussed.
Journal of CO2 Utilization 10 (2015) 67–77(Siddhartha Sengupta, Goutam Deo) 117
SURFACE AREA ANALYSIS
The surface areas of the Al2O3 and modified-Al2O3 supports and the supported Ni and Ni–Co catalysts were
determined and reported in Table 1.

118
UV–VIS SPECTRA ANALYSIS

UV–vis spectra under ambient conditions of: (a) 100Ni–Al, (b) 100Ni–Al– 2.5Ca, (c)
100Ni–Al–5Ca, (d) 100Ni–Al–2.5Mg and (e) 100Ni–Al–5Mg. The samples were calcined at
773 K.
119
Raman Spectra

Raman spectra obtained under ambient conditions of spent: (A) (a) 100Ni– Al, (b) 100Ni–Al–2.5Ca, (c) 100Ni–Al–5Ca,
(d) 100Ni–Al–2.5Mg and (e) 100Ni–Al– 5Mg and (B) (a) 75Ni25Co–Al, (b) 75Ni25Co–Al–2.5Ca, (c) 75Ni25Co–Al–5Ca,
(d) 75Ni25Co–Al–2.5Mg and (e) 75Ni25Co–Al–5Mg.

120
121
 CONCLUSION
The effect of modifying the support with CaO or MgO to improve the
catalytic activity of Al2O3 supported Ni and Ni–Co catalyst for the
DRM reaction was successfully investigated. The Al2O3 and modified-
Al2O3 supported Ni and Ni–Co catalysts were synthesized,
characterized and then tested for the DRM reaction. Characterization
of the supported Ni and Ni–Co catalysts revealed that modifying the
support with CaO or MgO had an effect on the reducibility, amount
and type of H2 and CO2 chemisorbed and the nature of the metal
crystallite. The reduction (%) of the supported Ni and Ni–Co catalysts
were not significantly affected by the presence of CaO. However, the
presence of MgO decreased the reduction (%) of the supported
catalysts. 122
Thank You

123