UCB008 - APPLIED CHEMISTRY

UCB008

Atomic Emission Spectroscopy

Principle
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 UCB008

Learning Outcomes
At the end of this session participants should be able to:

• Differentiate the principles associated with atomic emission and absorption

spectroscopy

Ranjana Prakash
 UCB008

Atomic Emission and

Atomic Absorption Spectroscopy

Ranjana Prakash
 UCB008

• In atomic emission and absorption spectroscopic technique sample solution

is aspirated into a flame that is hot enough to break the ions or molecules
into their atomic states.

• The concentration of the analyte in the flame can be measured by either

through absorption or emission of the radiation.

• The absorption mode is known as atomic absorption spectroscopy (AAS)

whereas emission mode as flame emission Spectroscopy (FES) or Atomic
emission spectroscopy.

Ranjana Prakash
 Na Li

UCB008

Ba Ca

Upper Diagram shows FES while the lower one shows AAS. In FES the flame provides
energy for atomization and excitation, but in AAS it provides only atomization .
Ranjana Prakash
 UCB008
Flame emission spectroscopy

• Flame emission spectroscopy (flame photometry) is a special area of emission

spectroscopy in which a flame is used to excite the atoms.

• For a few elements, such as the alkali metals Na and K, flame is hot enough
not only to produce ground state atoms, but to raise some of the atoms to an
excited electronic state.

• So Flame emission spectroscopy is used for the detection of alkali metals and
some of the alkaline earth metals.

Ranjana Prakash
 Flame emission spectroscopy
Sequence of events in flame
UCB008

Nebulization Desolvation Volatilization

Sample
solution

Spray Heat Dry aerosol Free

atoms
Heat

• Atomic absorption and atomic emission techniques involve introduction of

sample solution into a flame.
• The solution of the metal salt in question is sprayed into the flame.
• Solvent evaporates leaving the finely powdered salt.
• Vaporization of the salt.
• Conversion of ions into free gaseous atoms.
• Valence electron ( ) is raised to a higher energy state. Ranjana Prakash
 Flame emission spectroscopy
Nebulization – Conversion of sample solution in fine droplets – aerosol
formation which is aspirated to flame UCB008

Desolvation – Removal of solvent leaving dry sample

Sublimation- Transition of salt from solid to gaseous state

Atomization – Conversion of ions in to atoms

Excitation – Valence electrons of atoms get excited to higher energy state

Relaxation – Excited electrons relaxes from higher energy state to ground

state
Measurement – Wavelength and intensity of emitted radiations is measured

Ranjana Prakash
 UCB008
Flame emission spectroscopy

Principle
• Absorption of heat energy by ground state atom present in the flame
results in the excitation of valence electron of atoms.
• This valence electron comes back to ground state with the emission of
photon.
• Wave length and intensity of emitted photon helps in qualitative and
quantitative analysis of the sample.

Ranjana Prakash
The energy emitted when this electron drops down into a vacant lower
level is given off as radiant energy of a wavelength determined by the
UCB008
Planck-Einstein relationship:
E2-E1 = DE = hn = hc/l
l= hc/ E2-E1
l of the emitted radiation is characteristic of atoms of a particular
element from which it is emitted. It tells us about the elements which
are present in the flame
For example,
Na -------> Na* (energy from flame)
Na* -------> Na + hn (at 589 nm)

Ranjana Prakash
 UCB008
Intensity of radiant energy emitted when the atoms return to
the ground state is proportional to the concentration and is the
basis of flame emission spectroscopy.

The fraction of free atom that are thermally excited is governed

by boltzmann distribution law-

N*/N0 = A e-DE / kT

Ranjana Prakash
• According to Scheibe-Lomakin equation
I=kxC UCB008

Where :
I = Intensity of emitted light
C = Concentration of the element
k = Constant of proportionality

• The signal intensity from a flame is dependent on the flame temperature,

the rate of flow of liquid (sample solution) into the flame, the pressure and
rate of flow of fuel gases, and on many other variables which affect the
character of the flame or atomization of the sample.

• So flame temperature should be controlled. Flame temperature depends on

fuel, oxidant and fuel to oxidant ratio.

Ranjana Prakash
 Fuel Oxidant Flame temperature
(0C) UCB008

Propane Air 1900 oC

Propane Oxygen 2800 oC

Hydrogen Air 2100 oC

Hydrogen Oxygen 2800 oC

Acetylene Air 2200 0C

Acetylene Oxygen 3000 0C

Ranjana Prakash
 UCB008

In the next session…..

• Instrumentation of atomic emission
spectroscopy

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY
UCB008

Atomic Emission Spectroscopy

Instrumentation
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 UCB008

Learning Outcomes
At the end of this session participants should be able to:

• Associate the principle with instrumentation of flame photometry

Ranjana Prakash
 Instrumentation

UCB008
photodetector

prism
slit slit amplifier
O2 Fuel
Recorder

Sequence of events in flame

a) Sample solution is converted into a fine aerosol (nebulized) and

sprayed into the flame to desolvated, vaporized and atomized.
b) Excitation of atoms

Ranjana Prakash
 Instrumentation
UCB008

photodetector

prism
slit slit amplifier
O2 Fuel
Recorder

c) Excited atoms relax and emit radiation

d) The emitted radiant energy from flame emission is isolated
by a monochromator and detected by a photomultiplier.

Ranjana Prakash
 Instrumentation

UCB008
photodetector

prism
slit slit amplifier
O2 Fuel
Recorder
The emitted radiant energy from flame emission is isolated by a monochromator and
detected by a photomultiplier.
Burner- used for producing flame and burning sample solution
Flame – acts as an atomizer and emission intensity should be steady
Temp. of flame depends on-
1. Type of fuel, oxidant, fuel to oxidant rat
2. Type of solvent used for preparing sample solution
3. Amount of solvent entering in the flame 4.Type of burner used
Ranjana Prakash
 Instrumentation

UCB008
photodetector

prism
slit slit amplifier
O2 Fuel
Recorder

Ranjana Prakash
 UCB008

Applications
The technique is used to detect elements of group I and II of the periodic table
some elements can be detected visually by the colour in the flame

Ranjana Prakash
Ba Ca Na Li
UCB008

Ranjana Prakash
Qualitative Analysis

The technique is used to detect elements of group I and II of the periodic table UCB008

Within the flame, there are many more atoms in the ground state than in the excited

state.

For Zn, for instance, in a 2000K flame, there are 7.3x1015 atoms in the ground state

for every excited atom.

About 10 elements can be determined by conventional flame emission spectroscopy,

since a flame provides sufficient energy for only a few elements to have a significant

number of atoms in an excited state.

Ranjana Prakash
Two methods are used for preparing the calibration curve
- Standard addition method
UCB008
- Internal standard method
The alkali metals are elements with unoccupied atomic orbitals of low enough energy to
be sufficiently populated by a flame.
- Sodium and potassium ions play an important role in a number of biological systems
and their functions. Since these ions form few insoluble compounds and exhibit
essentially no acidic or basic properties, they cannot be determined readily by
conventional wet chemical techniques and are usually measured instrumentally.
- The usual techniques employed in determining these ions include atomic absorption
spectroscopy (AAS), atomic emission spectroscopy (AES).

Ranjana Prakash
Both these methods require that the sample exist as, or be converted to, an
UCB008
aqueous form.
Choosing between the individual methods is based primarily on the sensitivity,
speed and convenience of the method.
Detection limit for these ions is lowest by AAS but AES is simple, this method is
generally used for Sodium and potassium ions determination.

AES applications
Used for the analysis of biological fluids and tissues
Soil analysis
Natural and Industrial wastewater analysis

Ranjana Prakash
 UCB008

Limitations
• Less reliable than AAS
•Both the techniques does not provide any information about the molecular
structure of the compound
• Non radiating elements (C, H, halides) cannot be detected.

Ranjana Prakash
 UCB008

In the next session…..

• Categorization of various burners used in
atomic spectroscopy

Ranjana Prakash
 UCB008 - APPLIED CHEMISTRY
UCB008

Atomic Emission Spectroscopy

Instrumentation – Types of Burners
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 UCB008

Learning Outcomes
At the end of this session participants should be able to:

• Understand the working of various types of burners used for atomic

spectroscopic techiques

Ranjana Prakash
 UCB008

Types of burner
Mecker burner
Total consumption burner
Laminar flow burner

Ranjana Prakash
 Burners UCB008

Mecker Burner-

•Natural gas and oxygen are used as fuel and oxidant.

•Produces low temp, and low excitation energies.
• Used for alkali metals only.

Ranjana Prakash
Total consumption Burner

UCB008

• As soon as liquid sample is

drawn in the flame, oxygen
aspirates the sample solution.
• Desolvation
• Vapourisation
• Atomization
• Excitation
• Emission process
• Flame is noisy and turbulent but
can be adjusted to produce high
temperature by controlling fuel
to oxidant ratio.

Ranjana Prakash
Laminar-flow Burner (pre-mix)
• Sample, fuel and oxidant are thoroughly mixed
before reaching the burner opening and then
UCB008
enters in the flame
• Gases move in non-turbulent fashion – laminar
flow
• Only 5 percent of the sample in the form of
small droplets reaches to the flame and gets
decomposed
• Efficient atomization
• Larger droplets move out as condensate
• Flame is non-turbulent, noiseless and stable
• Disadvantages
– When sample contains two solvents, more volatile
solvent evaporates leaving sample in less volatile
solvent

– Therefore, smaller number of atoms would reach to

the flame and emission intensity reduces

Ranjana Prakash
 Comparison – Total Consumption & Pre-mix Burner
Total Consumption Burner Pre-mix Burner
Fuel, oxidant and sample meet together at Nebulized sample get mixed thouroughly UCB008
the base of the flame with fuel and oxidant then this mixture
goes in the flame
The flame is turbulent, noisy and non- The flame is non-turbulent, noiseless,
homogeneous stable and laminar flow
Entire sample that entered through the Only 5% of the sample in the form of
capiallary tube reaches to the flame and smaller droplets reaches to the flame and
get burnt regardless of dropsize burnt
Atomization is non homogenous Efficient and homogenous atomization
takes place
Preferred for flame photometry Perferred for atomic absorption
spectroscopy
Cost-effective Expansive

Ranjana Prakash
 UCB008

In the next session…..

• Principles associated with atomic absorption

spectroscopy

Ranjana Prakash
 UCB008

ATOMIC
ABSORPTION
SPECTROSCOPY
(AAS)

Ranjana Prakash
Atomic Absorption Spectroscopy
Principle UCB008

A sample solution containing known metal atoms, when introduced

into the flame, is irradiated with light of their own specific
wavelength will absorb light proportional to the density of atoms in
the flame.

Flame

Specific Wavelength

Thus, AAS measures Amount of Light Absorbed. 2

Ranjana Prakash
 Principle
UCB008

Absorption  density of atoms in flame

Amount of light absorbed  concentration of metallic species
Ability of atom to
Total amount of light absorbed = (πe2/mc) N.f  absorb at freq. n

No. of atom to absorb at freq. 

Total amount of light absorbed = constant x N x f

Therefore, absorption is independent of temperature and wavelength
Better than FES

Ranjana Prakash
 UCB008

Elements detectable by atomic absorption are highlighted in pink in this periodic table

Can be analyzed using AAS

Ranjana Prakash
 Atomic Absorption Spectroscopy UCB008

• Most powerful technique for the determination of trace metals in solution

• 70-80 elements can be detected
• Determination can be made in the presence of many other elements
• No specific sample preparation is required
• Wide application

Ranjana Prakash
 Differences between AAS and FES
FES
Excitation of atoms – emission of
photon and return to ground state.
Emission intensity is measured
Emission intensity  No. of
atoms in excited state
Emission intensity depends on
flame temperature
Beer’s law is not obeyed over
wide range of concentration
UCB008
AAS
Signal is obtained in presence and
absence of element and decrease in
intensity of signal obtained.
Absorption is measured
Absorption intensity  No. of
atoms in ground state
Absorption intensity does not
depend on flame temperature
Beer’s law is obeyed over a wide
range
Ranjana Prakash
 Advantages of AAS
UCB008
• AAS - specific – Atom of a particular element can absorb
radiation of their own wavelength – No spectral interference
• Much larger No. of atoms contribute in AAS signal so results are
more authentic
• Variation in flame temperature has less effect
• Absorption intensity does not depends on flame temperature
• Detection limit of AAS and FES is quite similar
• 70-80 elements can detected

Ranjana Prakash
 Disadvantages of AAS UCB008

• Different (HCL) lamp for each element is required

• Elements which for stable oxides eg. Al, Ti, W, Mo, do not give very good results

Ranjana Prakash
Instrumentation (premix or laminar flow
burner)
UCB008

INSTRUMENT FOR ATOMIC ABSORPTION : 1- Source of radiation(Hollow Cathode Lamp)

2- Chopper
3- Atomizer (Flame)
4- Monochromator (Prism)
5- Detector (Photo Cell)
6- Read out meter (Not in diagram)
Light separation and detection :
• AA spectrometers use monochromators and detectors for uv and visible light.
• The main purpose of the monochromator is to isolate the absorption line.
• Photo multiplier tubes are the most common detectors for AA spectroscopy Ranjana Prakash
• Absorption is measured by the difference in intensity of transmitted signal UCB008
in presence and absence of test element

• Production of atomic vapour- premix or laminar flow burner is used

Light separation and detection

• AA spectrometers use monochromators and detectors for uv and visible
light
• The main purpose of the monochromator is to isolate the absorption line
• Photo multiplier tubes are the most common detectors for AA
spectroscopy

Ranjana Prakash
Role of the Chopper
UCB008
• Breaks steady light coming from the hollow cathode lamp into intermittant light
• It gives a pulsating current to photodetector
• Absorption is measured without any interference
Role of the Flame
• Destroy any analyte ions and breakdown complexes
• Create atoms (the elemental form) of the element of interest
Fe0, Cu0, Zn0, etc.
Role of the Monochromator
• Isolate analytical lines photons passing through the flame
• Remove scattered light of other wavelengths from the flame
• In doing this, only a narrow spectral line impinges on the Photo multiplier tube.
Ranjana Prakash
 UCB008

Ranjana Prakash
 Sample Overview of AA
Compartment spectrometer.
UCB008

Light Source Detector

Ranjana Prakash
Job of the Hollow Cathode Lamp (Radiation source)

UCB008

Ranjana Prakash
Sequence of events in hollow-cathode lamp
► Ionization of rare gas atoms Ar + e-  Ar+ + 2e- UCB008

► Sputtering of cathode atoms: M(s) + Ar+  M(g) + Ar

► Excitation of metal atoms: M(g) + Ar+  M*(g) + Ar

► Light emission: M*(g)  M(g) + h

► The cathode contains the element that is analysed.
► Light emitted by hallow-cathode lamp has the same wave length as the
light absorbed by the analyte element
► Different lamp required for each element
Function of hollow cathode lamp
• Provide the analytical light line for the element of interest
• Provide a constant yet intense beam of that analytical line Ranjana Prakash
Hollow cathode lamp (Radiation source)
• It should emit stable intense radiation of the element to be determined, UCB008
resonance line of the element
• DC voltage is applied across anode and cathode
• Atom of the filler gas ionized at the anode and attracted by the cathode
• Inert gas ions strike the cathode and displace the surface metal atoms in
the inert gas
• Further collision of vaporized metal atoms with energetic inert gas ions
result in excited metal atoms emit the characteristic spectrum of the metal
used to construct the cathode
• Each HCL emits the spectrum of the metal used in the cathode
• Spectral lines emitted by the copper HCL can only be absorbed by the
copper atom present in the flame

Ranjana Prakash
 UCB008
Hollow cathode lamp (Radiation source)
• Hollow-cathode lamps are a type of discharge lamp that produce narrow
emission from atomic species. They get their name from the cup-shaped
cathode, which is made from the element of interest.
• The electric discharge ionizes rare gas atoms, which are accelerated into
the cathode and sputter metal atoms into the gas phase.
• Collisions with gas atoms or electrons excite the metal atoms to higher
energy levels, which decay to lower levels by emitting light.

Ranjana Prakash
 UCB008

• Atomic-absorption (AA) spectroscopy uses the absorption of light to

measure the concentration of gas-phase atoms. Since samples are usually
liquids or solids, the analyte atoms or ions must be vaporized in a flame.

• The atoms absorb ultraviolet or visible light and make transitions to higher
electronic energy levels.

Ranjana Prakash
 UCB008

• The analyte concentration is determined from the amount of absorption.

• Applying the Beer-Lambert law directly in AA spectroscopy is difficult due

to variations in the atomization efficiency from the sample matrix.

• Concentration measurements are usually determined from a working

curve after calibrating the instrument with standards of known
concentration.

Ranjana Prakash
 Calibration curve- Standard addition method
• Signal intensity of unknown sample X is taken
UCB008
• Series of solutions having standard sample of X are prepared and signal is obtained
by aspirating them in flame
• Plot is drawn between the signal intensity vs conc. of unknown X + Conc. of
standard X
• From the plot, the conc. of unknown can be determined from the intersection of the
curve with the conc. axis.

1.00 mL pipette

Fe:
0.05
mg mL-1
1 mL 2 mL 3 mL 4 mL 5 mL
50.00 mL volumetric flasks Ranjana Prakash
Standard Addition Curve
UCB008

o
X + 4X X + 6X
o
Signal X + 2X o
(Absorbance)
o
X+X
X

Conc. of unknown (x) Conc. of standard

Ranjana Prakash
Calibration curve- Standard addition method UCB008
• Signal intensity of unknown X is taken by aspirating sample solution to
flame
• Series of solutions having unknown X + varying amounts of standard X
are prepared and signal is obtained by aspirating them in flame
• Plot is drawn between the signal intensity against Conc. Of unknown X +
Conc. Of standard X
• From the plot, the conc. of unknown can be determined from the
intersection of the curve with the concentration axis.
• AAS compensates for any unexpected interfering material
present in the sample solution

Ranjana Prakash
Lead is extracted from a sample of blood and analyzed at 283 nm and gave an
absorbance of 0.340 in an AA spectrometer. Using the data provided, graph a
calibration curve and find the concentration of lead ions in the blood sample. UCB008

[Pb+2] (ppm) Absorbance Calculated Pb (II) concentraions (ppm) Absorbance

0.000 0.000 0.357 0.340
0.100 0.116
0.200
0.300
0.216
0.310
• The data provided in the problem
0.400
0.500
0.425
0.520
appears in the upper left hand
corner of this MS EXCEL
Lead (II) Calibration Curve worksheet.
0.600 • The graph is used to calculate the
0.500
best fit line.
y = 1.0505x

0.400
2
R = 0.9988 • The equation is then used to
calculate the concentration of Pb
Absorbance

0.300 (II) ions with an absorbance of

0.200
0.340.

0.100
• The result, 0.357 ppm, is
displayed above the graph.
0.000
0.000 0.100 0.200 0.300 0.400 0.500 0.600
[Pb+2] (ppm)

Ranjana Prakash
Advantages
AAS compensates for any unexpected interfering material present in the sample
solution UCB008

Ranjana Prakash
 SPECTROSCOPY UCB008

Atomic Spectroscopy: Introduction to atomic spectroscopy, atomic

absorption spectrophotometry and flame photometry.
Molecular Spectroscopy: Beer-Lambert`s Law, molecular
spectroscopy, principle, instrumentation and applications of UV-Vis
and IR spectroscopy.

Learning Outcomes: The students will be able to reflect on atomic and molecular
spectroscopy fundamentals like Beer`s law, flame photometry, atomic absorption
spectrophotometry, UV-Vis and IR.

School of Chemistry and Biochemistry

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - II

Spectroscopy - Introduction
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes

At the end of this session participants should be able to:

• Categorize molecular energy levels

• Illustrate types of molecular spectra

• Differentiate between atomic and molecular spectra

Ranjana Prakash
 SCBC-TIET

Molecular Energy Levels

• Emol = Eel + Evib + Erot + Etr
Eel >> Evib > Erot >> Etr (Etr is Negligible)
Emol = Eel + Evib + Erot
• Molecule – electronic, vibrational and rotational
energy levels
• Energy requirement for transition - Eel > Evib > Erot

Ranjana Prakash
 Energy levels in a molecule
∆Erotational < ∆Evibrational < ∆Eelectronic SCBC-TIET

J=3
J=2
Rotational transition
J=1 (microwave radiation
J=0 required)
E=2

E=1
Electronic transition
ν=2 (UV-Vis radiation required)
ν=1 Vibrational transition
ν=0 (IR radiation required)
E=0
Ranjana Prakash
Spectroscopy SCBC-TIET

• Deals with the transitions that a molecule or an atom undergoes

between its energy levels on the absorption of suitable radiation
determined by quantum mechanical selection rules
• Spectrum is a graph of the intensity of absorbed or emitted
radiation by sample verses frequency (n) or wavelength (λ)
• Spectroscopy involves characterization of spectrum of a sample
containing atoms or molecules
• The instrument used to obtain the spectrum of a compound is
called a Spectrometer/ Spectrophotometer
• Spectra of a compound is highly characteristic, thus,
spectroscopic techniques are used to determine the unknown
molecular structures

Ranjana Prakash
Spectroscopy SCBC-TIET

1. Absorption Spectroscopy:
• Absorption spectroscopy is based on the measurement of
absorption of electromagnetic radiation by atom or
molecule.
• UV (185 - 380 nm) / Visible (380 - 760 nm) Spectroscopy, IR
Spectroscopy
E2
p-hydroxybenzoic acid

DE = hn

E1

• lmax – Wavelength at which a molecule shows maximum absorbance

Ranjana Prakash
Spectroscopy SCBC-TIET

2. Emission Spectroscopy:
• Emitted radiations by atom or molecule are measured
• Fluorescence, phosphorescence , etc.
E2 Intensity

DE = hn

E1
n
Frequency

Ranjana Prakash
 Spectroscopy SCBC-TIET

Atomic spectroscopy Molecular spectroscopy

Absorption Emission
Atom when absorbs photon, its
electron jumps to higher energy
level and then returns back by
emission of photon having
energy equal to the energy
difference between two energy
levels
Absorption Emission
Molecule, when absorbs photon,
energy transitions take place
between molecular energy levels
Molecular spectrum

Atomic spectrum UV-Visible & IR Florescence

& Phosphorescence
Atomic absorption Flame emission

Ranjana Prakash
Types of Molecular spectra SCBC-TIET

Electronic or
UV-Vis. Spectra
(Electronic spectroscopy)

Vibrational spectra /
vibrational-Rotational
spectra / IR spectra
(IR/vibrational
spectroscopy)
Rotational Spectra
(Rotational/microwave
spectroscopy)
ESR spectra (ESR
spectroscopy)

NMR spectra
(NMR spectroscopy)

Ranjana Prakash
Atomic vs Molecular (Absorption) Spectrum - Difference SCBC-TIET

• Atomic spectrum - Line spectrum

• Molecular spectrum – Band spectrum

p-hydroxybenzoic acid

% Abs.
Absorbance (A)

Wavelength (l) l
Atomic Absorption Spectrum Molecular Absorption Spectrum
Ranjana Prakash
Atomic vs Molecular (Absorption) Spectrum - Difference
SCBC-TIET

• Atoms have only electronic energy

levels, whereas molecules have
electronic, vibrational and rotational
energy levels.
E=2
• Therefore, atoms have only electronic
transitions – Line spectrum
• However, in case of molecule,
electronic transitions are possible
Energy

from any vibrational or rotational

energy level of ground state to any
E=1 vibrational or rotational energy level
of excited state.
• Thus, vibrational and rotational
effects get super imposed over
electronic transition resulting in an
absorption band instead of
E=0 absorption lines
Ranjana Prakash
Spectroscopy
Molecule + EMR of suitable frequency SCBC-TIET

Absorption of radiation

l absorbed is Changes in elec., vibrat. & rot. energy

measured by levels of the molecules
spectrophotometer Highly
characteristic
of a molecule
% Abs.

Technique is
Used for
structure
l determination
Ranjana Prakash
 SCBC-TIET

In the next session…..

• Electronic transitions in the UV-visible spectroscopy

Ranjana Prakash
 UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - III

UV-Visible Spectroscopy – Electronic Transitions

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:

• Visualize and illustrate the types of electronic transitions

Ranjana Prakash
 UV-Visible Spectroscopy (or) SCBC-TIET

Electronic Spectroscopy

Antibonding
Molecular
ABMO
Orbital (ABMO)
hn
E Atomic Atomic UV-Vis
radiations
orbital orbital
Bonding BMO
Molecular
Excited State
Orbital (BMO)
Ground State

Ranjana Prakash
 UV-Visible Spectroscopy (or)
Electronic Spectroscopy SCBC-TIET

EM Radiations of UV–visible region Covalent bonds in organic molecule

+
compound having multiple bond

s bond p bond
Absorption of radiation

Transition of valence electron from
ground to excited state
s s* p p*
(bonding) (anti-bonding) (bonding) (anti-bonding)
• Compounds containing hetero atom have non-
bonding electrons
• A electronic transition is always from bonding
molecular orbital (BMO) to anti-bonding molecular
orbital (ABMO) Ranjana Prakash
 Types of Electronic Transitions
s* (bonding) SCBC-TIET

ss*
p* (bonding)
ns*
np* sp*
Energy

ps*
n (non-bonding)
pp*
p (bonding)

s (bonding)
Ranjana Prakash
 Energy Requirement for Electronic Transitions
SCBC-TIET

s-s* > n-s* ~ p-p* > n-p*

~150nm ~170-190nm 280nm

s-s* - saturated hydrocarbons e.g. ethane

n-s* - saturated compounds containing hetro atom having unshared pair

of electrons. e.g. saturated halides, alcohols, ethers, aldehydes, amines,
etc.

p-p* - compounds having double or triple bond and aromatics e.g.

butadiene, benzene, etc.

n-p* - unsaturated compounds containing hetro atom having unshared

pair of electrons e.g. carboxylic acids, aldehydes, ketones, etc.
Ranjana Prakash
 Electronic Transitions
SCBC-TIET

• An allowed electronic transition involves orbitals having same symmetry

e.g., ss* and pp* transitions.

• Allowed transitions have higher extinction coefficient (emax) values of

greater than 104.
• e.g., pp* transition of 1-3 butadiene absorbs at 270 nm (emax : 21000)

• Transitions between orbitals having different symmetry are classified as

symmetry forbidden transitions.
• e.g., s  p* and p  s* transitions are possible only theoretically .

• The forbidden transition namely n  p* have very low extinction coefficient

(emax) values.
• e.g., n  p* transition of carbonyl compounds, at 280 nm, has emax 15.
Ranjana Prakash
 Electronic Transitions

lmax
SCBC-TIET

Chromophore Example Excitation

(nm)
e
C=C Ethene pp* 171 15,000

C≡C 1-Hexyne pp* 180 10,000

np* 290 15
C=O Ethanal
pp* 180 10,000

np* 275 17
N=O Nitromethane
pp* 200 5,000

C-X Methyl bromide ns* 205 200

X=Br; X=I Methyl Iodide
ns* 255 360
Ranjana Prakash
 s-s* transitions SCBC-TIET

Far UV UV V I B G Y O R
s-s*
~150nm 100 200 380 760
nm nm nm nm
Vacuum ultraviolet
region

• s-s* requires photons of 150 nm.

• Conventional UV-visible spectrophotometer works in the range of
200 nm to 760 nm.
• This instrument cannot be used below 200 nm as oxygen in air
strongly absorbs in far UV region.
• Thus, s-s* transitions cannot be observed using conventional
UV-visible spectrophotometer.
Ranjana Prakash
 SCBC-TIET

s-s* transitions

• Analysis in far UV region requires vacuum ultraviolet

spectrophotometer which is cost intensive.
• Vacuum UV spectrophotometer is used mainly for determining
bond energies.
• This equipment is not used for organic structural determination.

Ranjana Prakash
 SCBC-TIET

In the next session…..

• Terms used in the UV-visible spectroscopy

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - IV

UV-Visible Spectroscopy – Terms

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:

• illustrate the terms used in UV-Visible Spectroscopy

Ranjana Prakash
 Chromophore SCBC-TIET

• Isolated covalently bonded groups that shows characteristic

absorption in UV-Visible region and even small structural changes
brings about change in wavelength of absorption.

or

• The functional groups containing multiple bonds capable of absorbing

radiations above 200 nm due to n-p* and p-p* transitions.

• Examples: C=C, C≡C, N=O, N=N, NO2, C=O, C=N, C≡N, C=C,C=S,
-CONH2, -COOH, etc

Ranjana Prakash
 SCBC-TIET

Chromophore

• Non-conjugated alkenes show an absorption below 200 nm and

therefore cannot be observed by conventional UV
spectrophotometer.

• If these transitions are to be observed, vacuum UV

spectrophotometer is required.

• Non-conjugated carbonyl group compound give a weak absorption

band in the 200 - 300 nm due to n-p* transition.

Ranjana Prakash
 Effect of conjugation on lmax SCBC-TIET

Compound lmax (nm)

H2C=CH2 180
217
O
220

220

255
Me
HO3S N N N
Me
460
Ranjana Prakash
 Effect of conjugation on lmax SCBC-TIET

CH2 CH3
H2C H3C

1,5 - hexadiene : λmax = 178 nm 2,4 - hexadiene : λmax = 227 nm

When double bonds are conjugated in a compound, λmax is shifted to

longer wavelength.

Ranjana Prakash
 Effect of conjugation on lmax
SCBC-TIET

O
C
H2C CH2 H3C CH3
Ethylene has λmax = 171 nm Acetone has λmax = 279 nm

Conjugation of C=C and carbonyl group shifts the λmax of both

groups to longer wavelength.
O

H2C C
CH3
Crotonaldehyde has λmax = 290 nm

Ranjana Prakash
 Auxochrome
SCBC-TIET

• The functional group with non-bonding electrons, that does not absorb
radiation in UV/visible region, when attached to a chromophore
increases the wavelength and intensity of absorption.
OR
• Any group which does not itself act as chromophore but its presence
brings about a shift of absorption bands towards red end of the
spectrum

Examples: -OH, -OR, -NH2 , -NHR, -NR2, -SH etc.

Ranjana Prakash
 Auxochrome SCBC-TIET

• Auxochrome when combined with chromophore, modifies the position

of absorption band relative to the parent chromophore
Chromophore + Auxochrome

New chromophore
having higher lmax

Examples: -OH, -OR, -NH2 , -NHR, -NR2, -SH etc.

• Auxochrome extends the conjugation of a chromophore by sharing

its lone pair of electrons
Ranjana Prakash
 Auxochrome SCBC-TIET

e.g. Benzene λmax = 255 nm

OH

Phenol λmax = 270 nm

NH2

Aniline λmax = 280 nm

Ranjana Prakash
 SCBC-TIET

In the next session…..

• Absorption and Intensity Shifts in UV-Visible Spectroscopy

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - V

UV-Visible Spectroscopy – Absorption and Intensity Shifts

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes

At the end of this session participants should be able to:

• Distinguish between various shifts and effects associated with

UV-visible spectroscopy

Ranjana Prakash
 Shifts and Effects SCBC-TIET

Hyperchromic effect
Absorbance ( A )

Red shift
Blue shift
(Bathochromic
(Hypsochromic
shift)
shift)

Hypochromic effect

λmax
Wavelength ( λ ) Ranjana Prakash
 SCBC-TIET

Bathochromic Shift (Red Shift)

• When absorption maximum (emax) is shifted towards longer wavelength,

it is known as bathochromic shift or red shift.

• The effect is due to presence of an auxochrome or by the change of

solvent.

• For example, an auxochrome group like –OH, -OCH3 causes

absorption of compound at longer wavelength.

Ranjana Prakash
 Bathochromic Shift (Red Shift)
SCBC-TIET

• p-nitrophenol, in alkaline medium, shows red shift, because negatively

charged oxygen delocalizes more effectively than the unshared pair of
electron.
- -
O + O O + O
N N

-
OH

A lk a lin e
m e d iu m -
OH O

λmax = 255 nm λmax = 265 nm

p-nitrophenol p-nitro-phenoxide

Ranjana Prakash
 Hypsochromic Shift (Blue Shift) SCBC-TIET

• When absorption maximum (emax) is shifted towards shorter

wavelength, it is known as Hypsochromic shift or blue shift.
• The effect is due to removal of conjugation or by the change of
solvent.
• Aniline shows blue shift in acidic medium because it loses
conjugation.
+ -
NH2 + NH 3 Cl
H
A c id ic Conjugation decreases
due to protonation
m e d iu m

λmax = 280 nm λmax = 265 nm

Aniline Anilinium chloride
Ranjana Prakash
 Hyperchromic effect SCBC-TIET

• When intensity of absorption maximum (emax) of a compound

increases, it is known as hyperchromic effect.

N
A=ecx N CH3
λmax = 257 nm λmax = 260 nm
ε = 2750 ε = 3560
Pyridine 2-methyl pyridine

• Inclusion of an auxochrome also increases intensity of absorption.

Sharma YR, Elementary Organic Spectroscopy, S. Chand & Co. Ranjana Prakash
 Hypochromic effect SCBC-TIET

• When intensity of absorption maximum (emax) of a compound decreases,

it is known as hypochromic effect.

CH3
e = 19000 e = 10250
λmax = 250 nm λmax = 237 nm
Naphthalene 2-methyl naphthalene

• emax decreases due to the distortion caused by the methyl group.

Sharma YR, Elementary Organic Spectroscopy, S. Chand & Co.

Ranjana Prakash
 SCBC-TIET

In the next session…..

• Understand the effect of conjugation on lmax
• Colour in organic compounds

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - VI

UV-Visible Spectroscopy – lmax and Conjugation

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes

At the end of this session participants should be able to:

• Understand the effect of conjugation on lmax

Ranjana Prakash
 SCBC-TIET

How conjugation causes bathochromic shift….

• More the number of double bonds in conjugation, longer wavelength
photon is required for transition.
• Thus, energy requirement for electronic transition decreases.
• Compounds having > 8 double bonds in conjugation will appear
coloured to human eye.
• Energy requirement for pp* transitions decreases as the
conjugation increases and can reach the visible region of the spectrum
which make the compounds coloured.

Ranjana Prakash
 SCBC-TIET

How conjugation causes bathochromic shift….

• An electronic transition is from bonding molecular orbital to anti-

bonding molecular orbital.

• Energetically favored electron promotion will be from the highest

occupied molecular orbital (HOMO) to the lowest unoccupied
molecular orbital (LUMO).

• This results in excited state.

Ranjana Prakash
 Relative energies of orbitals - electronic spectroscopy
SCBC-TIET

s* s*

p4* Lowest
Unoccupied
p* p* Molecular Orbital
p3* (LUMO)
Energy

n n Highest Occupied
Molecular Orbital
p2 (HOMO)
p p
p1

s s
C-C C=C C=O C=C-C=C
s-s* p-p* p-p*
n-p*
n-s* p - p*
2 3
Kemp, W., Organic Spectroscopy, Palgrave Publ. Ranjana Prakash
Effect of conjugation on relative energies of molecular orbitals
SCBC-TIET

Ranjana Prakash
 How conjugation causes bathochromic shift….
SCBC-TIET

• When two double bonds are conjugated, the four p-

atomic orbitals combine to generate four p -molecular
orbitals (two are bonding and two are antibonding).
• When two double bonds are in conjugation the
energy level of HOMO is raised and that of LUMO is
lowered.

• The energetically most favorable p  p* excitation

occurs from the highest energy bonding pi-orbital
(HOMO – p2) to the lowest energy antibonding pi-
orbital (LUMO – p3*).
• In a similar manner, the three double bonds of a
conjugated triene create six p - molecular orbitals,
half bonding and half antibonding.
Ranjana Prakash
 SCBC-TIET

In the next session…..

• Development of colour in organic compounds

Ranjana Prakash
 UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - VII

UV-Visible Spectroscopy – Colours in organic compounds

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:

• Understand the origin of colours in organic compounds

Ranjana Prakash
 Colour in organic compounds
SCBC-TIET

• When white light passes through a coloured substance, a specific portion of the certain
wavelengths is absorbed.
• The remaining portion of light acquires
620 nm
the complementary colour to the
wavelength(s) absorbed.

• Thus, absorption of 430-490 nm light

gives orange colour to a substance, and
absorption of 500-540 nm light makes it
red.

490 nm • Green colour will develop if the

absorption at the start (400 nm) or end
• Complementary colours - (800 nm) of the visible region.
diametrically opposite to each
other. Ranjana Prakash
 Absorbed Light Observed colour SCBC-TIET

Wavelength (nm) Corresponding colour (transmitted)

400 Violet Yellow-green
425 Indigo-blue Yellow
450 Blue Orange
490 Blue-green Red
510 Green Purple
530 Yellow-green Violet
550 Yellow Indigo-blue
590 Orange Blue
640 Red Blue-green
730 Purple Green

Pretsch, Clerc, Seibl, Simon: Spectral Structure Determination of Organic

Compounds, Springer Science Ranjana Prakash
 Colour in organic compounds
SCBC-TIET

Number of conjugated
double bonds

5 colourless
Naphthalene

7 colourless

Anthracene

9 orange

Tetracene

Ranjana Prakash
 Colour in phenolphthalein
SCBC-TIET

Colourless 2 1 Pink
2 1
4 4 lmax = 552 nm
3 6
3
5

7 8
colourless pink

Ranjana Prakash
 Colours in nature SCBC-TIET

620 nm

Lycopene : 443-502 nm
446
475
421

Lutein : 421-475 nm

452

490 nm

b-carotene : 452 nm

Ranjana Prakash
 SCBC-TIET

In the next session…..

• UV-visible spectroscopy – Instrumentation

Ranjana Prakash
 UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - VIII

UV-Visible Spectroscopy – Instrumentation and Analysis

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:

• distinguish various functional components of a UV-visible spectrophotometer

• differentiate between single-beam and double-beam UV-visible spectrophotometer

Ranjana Prakash
 UV-Vis Spectrophotometer SCBC-TIET

• Light source
– Deuterium lamp – UV radiation
– Tungsten filament lamp – Visible radiation
• Sample containers
– Cuvettes
• Plastic
• Glass
• Quartz

Ranjana Prakash
 Sample Handling SCBC-TIET

Solvent - Should not absorb in the region under investigation

Solvent
1 mg sample 100ml

Sample solution
1 cm

Solvents – water, ethanol, n-hexane, cyclo-hexane, benzene, methyl

alcohol, diethyl ether etc.

Ranjana Prakash
 Sample Handling SCBC-TIET

Sample solution
1 cm

• Cuvettes are used for keeping the sample solution in the instrument.
• Thickness of sample solution in cuvette – 1cm
• Cuvette material should not absorb in the region under investigation.
• Glass/plastic – used only for visible region as they absorb UV radiation
• Quartz – used for UV-visible region

Ranjana Prakash
 Monochromator SCBC-TIET

• Monochromator is used to isolate the required wavelength from

polychromatic radiations
• Entrance slit: Sends a uni-directional beam to prism
• Prism: Disperses the radiations into constituent wavelengths
• Exit slit: Sends monochromatic beam to sample solution

Monochromator
Entrance slit Exit slit

Ranjana Prakash
 Detector
SCBC-TIET

• Detector detects the radiation received and converts them into electrical
signal.

Amplifier
• The signal received from the detector is amplified by the amplifier and sent
to read-out device.

Output
• The read-out device gives the absorbance value at different wavelengths
which can be plotted as an absorbance vs wavelength graph.

Ranjana Prakash
 UV-Vis Spectrophotometer (Single Beam)
SCBC-TIET

Visible
light
Source
(380-760 nm)

Output
Monochromator Blank
Sample
UV light Entrance Exit Cuvette
inSolution Detector Amplifier
Source slit in Cuvette
slit
(200 – 380 nm)

Ranjana Prakash
UV-Vis Spectrophotometer
(Double Beam) SCBC-TIET

Ranjana Prakash
UV-Vis Spectrophotometer
SCBC-TIET

Single Beam

Double Beam

Ranjana Prakash
Single Beam SCBC-TIET

• Solvent is taken as blank solution for which

absorbance is measured and absorbance value is set Io
to Zero and transmitted intensity is considered as Io.

• Absorbance of sample solution is measured, and if

sample absorbs radiations, then transmitted intensity I Io I
is less than Io.
Absorbance = log Io / I

• Instrument gives output graph which is plot of

wavelength of entire region vs absorbance of radiation
at each wavelength of the region which is termed as
Spectrum.
Ranjana Prakash
• Double Beam
SCBC-TIET

• In Double beam spectrophotometer, each absorbance measurement of

solution of sample is accompanied by simultaneous measurement of the
pure solvent
• Instrument is capable in comparing the intensities of the two beams at each
wavelength of the region

p-hydroxybenzoic acid
• In UV-visible spectrophotometers, The ratio between reference
(blank) beam and sample beam intensities (Io / I), i.e., ratio
recording is carried out.
• Absorbance = log Io / I
• A = ecx – Beer’s Law, where concentration (c) and thickness of
sample solution (x) are constant for a given sample lmax Ranjana Prakash
UV-visible spectrophotometers in our laboratories..
SCBC-TIET

…at School of Chemistry and Biochemistry

…at School of Energy and Environment

Ranjana Prakash
 SCBC-TIET

In the next session…..

• Qualitative and quantitative analysis
• Applications of UV-visible spectroscopy

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Molecular Spectroscopy Series

Lecture - IX

UV-Visible Spectroscopy –Analysis and Applications

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:
• Understand the analytical procedure of qualitative and
quantitative analysis using UV-visible spectrophotometer
• Gain knowledge on applications of UV-visible spectroscopy

Ranjana Prakash
Qualitative analysis using UV-visible spectrophotometer SCBC-TIET

• lmax of organic compounds can be determined using UV-visible spectra

• This value can be used to detect the extent of conjugation
• … and absence/presence of the chromophore in an organic compound.

lmax
Ranjana Prakash
Applications SCBC-TIET

• Qualitative Analysis:
• Extent of conjugation in polyenes can be estimated.
• Shift in the absorption, due to addition of unsaturation in a compound,
towards longer wavelength can be detected.
• Presence or absence of a chromophore can be detected.
• If UV-visible spectrum of a compound has no peaks above 200 nm, it
shows the absence of conjugation, carbonyl group and aromatic ring.
• It is used for characterizing aromatic compounds and conjugated
olefins.
• Distinction between conjugated and non-conjugated compounds
(CH3)2C=CH-CO-CH3 CH2=C(CH3)-CH2-CO-CH3
A B Ranjana Prakash
Applications SCBC-TIET

• Qualitative Analysis:
• Geometric isomers can be distinguished

H
C=C C=C
H H
H
cis-stilbene trans-stilbene
lmax : 283 nm lmax : 295 nm

• Trans isomer always absorb at higher wavelength

Ranjana Prakash
Applications SCBC-TIET

• Qualitative Analysis:
• Identification of a compound in different solvents- If structure of a
compound changes in different solvents, the change can be detected
by uv-vis spectra.
CCl3CHO.H2O

Hexane Water

290nm No absorption at 290nm (absence of C=O)

CCl3C=O.H2O CCl3C OH
OH
H H

Sharma YR, 2010, Elementary Organic Spectroscopy, S.Chand & Co.

Ranjana Prakash
Quantitative analysis of an unknown sample solution using UV- SCBC-TIET

visible spectrophotometer
• Series of standard solutions having known concentration of analyte X are
prepared.
• One of these solutions is analyzed by the spectrophotometer and its lmax
is determined.
• Absorbance of all the standard solutions are analyzed at the determined
lmax.
• A calibration curve (absorbance vs concentration) is prepared for the
standard samples.
• Signal intensity of unknown sample of analyte X is taken.
• The concentration of the analyte X is then determined based on the
absorbance value from the calibration curve.

Ranjana Prakash
Quantitative analysis of an unknown sample solution using UV-
visible spectrophotometer SCBC-TIET

Step 1: Series of standard solutions having known concentration of analyte

X are prepared.

Ranjana Prakash
Quantitative analysis of an unknown sample solution using UV-
visible spectrophotometer SCBC-TIET

Step 2: Solvent is taken as blank solution for which absorbance is measured

and absorbance value is set to Zero.

Io

Blank Solution

Ranjana Prakash
Quantitative analysis of an unknown sample solution using UV-
visible spectrophotometer SCBC-TIET

Step 3: One of the standard solutions is analyzed by the UV-visible

spectrophotometer and its lmax is determined.

lmax

Instrument is calibrated with blank, periodically, when wavelength is changed,

in case of single beam spectrophotometer.
Ranjana Prakash
Quantitative analysis of an unknown sample solution using UV-
visible spectrophotometer SCBC-TIET

Step 4: Absorbance of all the standard solutions are analyzed at the

determined lmax.
Step 5: A calibration curve (absorbance vs concentration) is prepared by
analyzing the standard samples.

3.0
2.5
Absorbance

2.0
1.5
1.0
0.5
0
0 1 2 3 4 5
Concentration (mg/ml)
Ranjana Prakash
Quantitative analysis of an unknown sample solution using UV-
visible spectrophotometer
SCBC-TIET

Step 6: Absorbance value of the solution of analyte X (unknown) is

determined at the same lmax.
Step 7: The concentration of the analyte X is then determined based on the
absorbance value from the calibration curve by extrapolating
towards concentration in X axis.
3.0 - concentration of analyte X
2.5 - e.g., ~3.66 mg/ml in this case
Absorbance

2.0
1.5
1.0
0.5
0
0 1 2 3 4 5
Concentration (mg/ml) Ranjana Prakash
Applications SCBC-TIET

• Quantitative Analysis:
• It can be used to find out molar concentration of the solute under study.

3.0
2.5
Absorbance

2.0
1.5
1.0
0.5
0
0 1 2 3 4 5
Concentration (mg/ml)

Ranjana Prakash
 SCBC-TIET

With this session…..

• We complete the understanding of the fundamental aspects associated
with UV-visible spectroscopy

From the next session…..

• We will be exploring the principle, instrumentation and applications of
Infra-red spectroscopy

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Infra-red Spectroscopy Series

Lecture - I

IR Spectroscopy - Introduction
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:
• Identify an IR spectrum of a compound
• Understand relationship between vibrational transitions and IR
spectroscopy

Ranjana Prakash
Microwave Spectroscopy SCBC-TIET

• Rotational energy level transitions of the gaseous molecules on the

absorption of radiations falling in microwave region.
• Molecule should necessarily have permanent dipole moment.
• Ex. HCl, CO, H2O vapour, NO, etc.

Ranjana Prakash
Infra-red spectroscopy
• The light our eyes see is a small part of a broad spectrum of SCBC-TIET

electromagnetic radiation.
• On the immediate high energy side of the visible spectrum lies the
ultraviolet, and on the low energy side is the infrared.

Ranjana Prakash
Infra-red spectroscopy
SCBC-TIET

• The portion of the infrared region most useful for analysis of organic
compounds is not immediately adjacent to the visible spectrum

IR region

Near IR IR Far IR
0.8-2.5 m 2.5-15 m 15-200 m

• Photon energies associated with infrared region are not large enough to
excite electrons, but may induce vibrational excitation of covalently
bonded atoms and groups.

Ranjana Prakash
Infra-red spectroscopy
SCBC-TIET

• Absorption of IR radiations can be expressed in terms of wavelength

or wave number ( ) –preferred

in cm−1

for ex. l = 15 m = 15 x 10-4 cm

in cm-1 =

0.8 m 2.5 m l 15 m 200 m

NIR IR FIR
12,500 4000 667 50 cm-1
Ranjana Prakash
 Infra-red spectroscopy
SCBC-TIET

0.8 m 2.5 m l 15 m 200 m

NIR IR FIR
12,500 4000 667 50 cm-1

Absorbance (A)
Change in A = log (1 / T)
intensity
Transmittance (T) -preferred

• Infra-red spectra are plotted as percent transmittance (T) vs wave

number ( )

Ranjana Prakash
 Infra-red spectroscopy
SCBC-TIET

Source: https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/InfraRed/infrared.htm
Ranjana Prakash
IR Spectra-vibrational transitions
SCBC-TIET
• Vibrational rotational spectroscopy - Vibrational energy change is accompanied
with large number of rotational energy changes.
• Vibrational energy levels - Quantized
• v – Vibrational quantum number i.e., 0,1,2,3,…….
n - Vibrational frequency
ଵ
 ௏ v
ଶ

 Lowest Vibrational level -v = 0,
ଵ ଵ
 ௏ = : Zero point energy
ଶ ଶ

E 1= E2 = E3 =
E2- E1 = E1 – E0 =
Ranjana Prakash
IR Spectra-vibrational transitions
SCBC-TIET

• Absorption of IR radiation equal to the energy difference between two

vibrational levels cause a vibrational transition
• Transition from ground state (v = 0) to first excited state ( v = 1) absorbs IR
radiations strongly and gives rise to intense band called the Fundamental
Band.
-

• Transition from ground state (v = 0) to second excited state ( v = 2) gives rise

to weak band called the Overtone.

-
Ranjana Prakash
IR Spectra-vibrational transitions SCBC-TIET

• All the vibrational energy levels are equally spaced by an

spacing of
v

 v - Operative part

Ranjana Prakash
 SCBC-TIET

In the next session…..

• Molecular vibrations and Hooke’s law

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Infra-red Spectroscopy Series

Lecture - II

IR Spectroscopy – Molecular Vibrations

by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:
• Distinguish various types of vibrational motions in a molecule
• Calculate vibrational frequency using Hooke’s law

Ranjana Prakash
• Molecular Vibrations SCBC-TIET

• The covalent bonds in molecules are not rigid sticks or rods, such as
found in molecular model kits, but are more like stiff springs that can be
stretched and contracted.
• Molecules - consisting of balls (atoms) of different sizes tied with
springs (bonds) of varying strength.

Atom
Chemical bond

Ranjana Prakash
Molecular Vibrations
SCBC-TIET
• Infrared radiation induces vibrational transitions in covalently bonded atoms or
molecules.
• Diatomic molecules having covalent bond can be assumed as spring holding together
two atoms.
Equilibrium bond length

Compressed covalent bond

Stretched covalent bond

Specific bonds respond to (absorb) specific frequencies of the photons of IR region

Ranjana Prakash
 Molecular Vibrations
SCBC-TIET

Stretching vibrations Bending vibrations

(Distance between atoms changes) (Bond angle changes)

atom

Chemical bond
Ranjana Prakash
 Molecular Vibrations SCBC-TIET

Stretching vibrations Bending vibrations

(Distance between atoms changes) (Bond angle changes)

Inplane Out of plane

Symmetric Asymmetric
stretching stretching Scissoring Rocking Wagging Twisting

 Asym   Sym   Bending

Animation courtesy: https://en.wikipedia.org/wiki/Molecular_vibration
Ranjana Prakash
Molecular Vibrations SCBC-TIET

• In addition to the facile rotation of groups about single bonds, molecules

experience a wide variety of vibrational motions, characteristic of their
component atoms.
• Consequently, virtually all organic compounds will absorb infrared radiation
that corresponds in energy to these vibrations.
• Infrared spectrometers permit chemists to obtain absorption spectra of
compounds that are a unique reflection of their molecular structure.
• Frequency of absorption depends on
1.Relative masses of the atoms
2.Force constant – Bond strength
3.Arrangement of atom within the molecule

Ranjana Prakash
Hooke’s Law and Molecular Vibrations SCBC-TIET

Frequency of absorption - Calculation – Hooke’s Law

Where:  = oscillating frequency (Hz)

Hz k = force constant
m = reduced mass of the atoms

cm-1
Where: = Wave number (cm-1)
c = speed of light (3 x 1010 cm/s)

Where: M1 & M2 = masses of atom in kg

IR spectra is recorded as transmittance vs wave number (cm-1)

Ranjana Prakash
Hooke’s Law and Molecular Vibrations SCBC-TIET

Hz

If Bond strength increases vibrational frequency

or increases
Reduced mass (m) decreases

If Bond Strength decreases vibrational frequency

or decreases
Reduced mass (m) increases

Ranjana Prakash
 SCBC-TIET

In the next session……

• Principles underlying IR spectroscopy

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Infra-red Spectroscopy Series

Lecture - III

IR Spectroscopy - Principles
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:
• Understand the principles of IR spectroscopy

Ranjana Prakash
Principles of IR spectroscopy
SCBC-TIET

Criteria for a molecule to absorb IR radiation

• Correct wavelength of radiation
• A molecule will absorb IR radiation when natural frequency of vibrations of some
part of a molecule is same as that of the frequency of incident radiation.
• Change in dipole moment
• A molecule is said to have an electric dipole when there is a slight positive and a
slight negative charge on its component atoms.
• Vibrational motion of the molecule should be accompanied by change in
(fluctuating) dipole moment i.e., dipole moment must change during the
vibration
• With change in dipole moment, charge distribution of the molecule changes & it
is the net change in charge distribution produced during stretching or bending
which makes possible interaction between the molecule and IR radiations
Ranjana Prakash
Principles of IR spectroscopy SCBC-TIET

• Type of vibration
• Stretching vibrations: higher frequency
• Bending vibrations: lower frequency
• Masses of the atoms in a bond [Atomic weight increases, Frequency
decreases]
• Strength of the bond or bond order [Bond order increases, Frequency
increases]
• Vibrational transition that causes a significant change in the dipole
moment of a chemical bond leads to strong absorption bands – IR active
transitions, for example, Carbonyl gp
• Vibrations that result in no change in dipole moment does not give rise to
any absorption band – IR inactive, for example, Homonuclear diatomic
molecules
Ranjana Prakash
Vibrational modes SCBC-TIET

• For a molecule, number of possible vibrational modes depends on its

vibrational degrees of freedom (DoF)
• Each atom has 3 DoF corresponding to the 3 cartesian coordinates (X, Y, Z)
• When atoms combine to form a molecule, no DoF is lost
• Total of DoF of a molecule = 3n (n  number of atoms)
• 3n Degree of Freedom = translational + rotational + vibrational
• A molecule, always, has 3 translational DoF
• A non-linear molecule has 3 rotational DoF but a linear molecule has only 2
rotational DoF, because rotation of linear molecule about its axis of linearity
does not bring any change in the position of the atoms.
Ranjana Prakash
Vibrational modes SCBC-TIET

• 3n Degree of Freedom = translational + rotational + vibrational

• Vibrational DoF for linear molecule = 3 + 2 + (3n-5)

• Vibrational DoF for non-linear molecule= 3 + 3 + (3n-6)

Molecule Number of Total DoF Translational Rotational Vibrational

atoms DoF DoF DoF
Carbon dioxide 3 3n = 3 x 3 = 9 3 2 9–5=4

H2O 3 3n = 3 x 3 = 9 3 3 9–6 =3

Benzene 12 3n = 3 x 12 = 36 3 3 36 – 6 = 30

Toulene 15 3n = 3 x 15 = 45 3 3 45 – 6 = 39

Acetylene 4 3n = 3 x 4 = 12 3 2 12 – 5 = 7

Ranjana Prakash
Principle of IR spectroscopy SCBC-TIET

• Diatomic molecules
• For a diatomic molecule, 3n = 3 x 2 = 6
• Vibrational DoF for diatomic molecule = 3n – 5 = 3 x 2 – 5 = 1

• Thus, for a diatomic molecule – only stretching vibration is possible

Diatomic Molecule

Homonuclear Heteronuclear
H2, N2, O2 CO, NO, HCl
IR inactive IR active

Ranjana Prakash
Principle of IR spectroscopy
• Triatomic molecules SCBC-TIET

• For a molecule, number of possible vibrational modes depends on its

vibrational degrees of freedom (DoF)
• Vibrational DoF increases with increase in number of atoms in a molecule
• Out of these all possible vibrational modes, only those which causes
fluctuating dipole moment in molecule give rise to absorption in IR region
• If a molecule has center of symmetry, vibrations which are centro-symmetric
are IR inactive
• Out of all possible vibrational modes, even if only one mode is active in IR
region, molecule is said to be IR active
• Therefore, all triatomic and polyatomic molecules are IR active
• Except, homonuclear diatomics, all other molecules are IR active
Ranjana Prakash
CO2 – Vibrational Modes
SCBC-TIET
• Asymmetric Stretch

• Symmetric Stretch (not IR active)

• Vertical Bend

• Horizontal Bend
(A degenerate mode with same motion as above but
rotated by 90o)

Ranjana Prakash
 CO2 – IR Spectrum
SCBC-TIET

Asymmetric stretch Bending

CO2 – IR spectrum

(667 cm-1)

(2349 cm-1)

• Number of possible vibrational modes = 3n – 5 = 3 x 3 – 5 = 4

• Number of IR active vibrational modes = 3 (symmetric stretching – IR inactive)
• Number of bands in IR spectrum = 2 (both bending modes are degenerate)
Ranjana Prakash
 H2O – Vibrational Modes
SCBC-TIET

O O O

H H H H H H
Symmetric Asymmetric Bending
• All three modes of vibration in H2O are not centrosymmetric and hence IR active
• Therefore, water cannot be used as solvent, in case of IR spectroscopy

Source: http://www1.lsbu.ac.uk/water/water_vibrational_ spectrum.html

Ranjana Prakash
 SCBC-TIET

In the next session…..

• Instrumentation associated with IR spectroscopy

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Infra-red Spectroscopy Series

Lecture - IV

IR Spectroscopy - Instrumentation
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:
• Relate principles of IR with its instrumentation

Ranjana Prakash
Instrumentation - IR SCBC-TIET

• IR Source
• …should give a continuous and high radiant energy output over entire IR
region
Globar – Rod of silicon carbide, electrically heated – 12000C

Nernst Glower – Mixture of oxides of Yt, Zr and Th molded in the shape

of rod – electrically heated upto 19000C

Ranjana Prakash
 SCBC-TIET

Instrumentation - IR
• Sample Handling
• For gases and liquid samples
• Commercial cells made up of alkali metal halide (NaCl) are used
• NaCl is hygroscopic and should be protected from moisture

Ranjana Prakash
Instrumentation - IR
SCBC-TIET
• Sample Handling
• For solid samples – KBr Disc technique
• Can be analyzed as alkali metal halide (KBr) mixture
• Sample should be free from moisture
Solid sample (0.1-0.3%) + KBr (powdered)  pressed in dye (under
vacuum) 8 ton pressure for 5 mins; 10 ton pressure for 1 min

13 mm disc

IR examination

• A pure KBr disc should be used as reference material

Ranjana Prakash
 Instrumentation - IR SCBC-TIET

Less Intense (I) A = log I0 / I

Globar T = I / I0
A = log 1 / T

Mono-
chrom
-ator

Nernst Glower

No change (I0)

Ranjana Prakash
Instrumentation - IR
SCBC-TIET

IR Source A = log I0 / I
T = I / I0
Beam Splitter A = log 1 / T

Beam Beam

Sample Reference

Sample beam Reference beam

Less Intense (I) No change (I0)

Ranjana Prakash
 SCBC-TIET

In the next session…..

• Interpretation of IR spectra

Ranjana Prakash
UCB008 - APPLIED CHEMISTRY SCBC-TIET

Infra-red Spectroscopy Series

Lecture - V

IR Spectroscopy - Interpretation
by
Prof. Ranjana Prakash
School of Chemistry and Biochemistry
Thapar Institute of Engineering and Technology
Patiala -147004, India
Ranjana Prakash
 SCBC-TIET

Learning Outcomes
At the end of this session participants should be able to:
• Interpret an IR spectra

Ranjana Prakash
Interpretation of IR spectra
SCBC-TIET

/
1 𝑘
𝑣̅ =
2𝜋𝑐 𝜇
cm-1

Functional group region

Fingerprint region

Source: https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/InfraRed/infrared.htm
Ranjana Prakash
Interpretation of IR spectra
SCBC-TIET

Source: https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/InfraRed/infrared.htm
Ranjana Prakash
Interpretation of IR spectra
Functional group region Fingerprint region SCBC-TIET

/
1 𝑘
𝑣̅ =
2𝜋𝑐 𝜇
cm-1

Ranjana Prakash
 SCBC-TIET
Interpretation of IR spectra

• The complexity of infrared spectra in the 1450 to 600 cm-1 region makes it
difficult to assign all the absorption bands, and because of the unique
patterns found there, it is often called the fingerprint region.
• Absorption bands in the 4000 to 1450 cm-1 region are usually due to
stretching vibrations of diatomic units, and this is sometimes called the
group frequency region.
• Group frequency region is also known as Peak Identification Area or
Functional Group region.

Ranjana Prakash
Interpretation of IR spectra SCBC-TIET

• For example:
CH3OH C2H5OH CH3CH2CH2OH

• If the two samples have same functional group, they show similar
absorption above 1450 cm-1, but their spectra will differ in fingerprint
region.

• If two samples are giving same spectra in fingerprint region, then

they possess same structures or they are enantiomers.

Ranjana Prakash
Interpretation of IR spectra
SCBC-TIET
Some General Concepts
• Stretching frequencies are higher than corresponding bending
frequencies. (It is easier to bend a bond than to stretch or compress it)
• Triple bonds have higher stretching frequencies than corresponding
double bonds, which in turn have higher frequencies than single bonds
• Bonds to hydrogen have higher stretching frequencies than those to
heavier atoms
• Since most organic compounds have C-H bonds, a useful rule is that
absorption in the 2850 to 3000 cm-1 is due to sp3 C-H stretching;
whereas, absorption above 3000 cm-1 is from sp2 C-H stretching or sp
C-H stretching if it is near 3300 cm-1

Ranjana Prakash
Interpretation of IR spectra
SCBC-TIET
Some General Concepts

• Presence of bands above 1500 cm-1 indicates the presence of specific

functional group in a compound
• Characteristic wave number of absorption in IR spectra for different
groups present in various classes of organic compounds are already
been identified and reported in literature
• Group frequency region is also known as Peak Identification Area or
Functional Group region.

Ranjana Prakash
Interpretation of IR spectra
Carbon-Carbon Bonds SCBC-TIET

• Increasing bond order leads to higher frequencies:

• C-C 1200 cm-1 (fingerprint region)
• C=C 1600 - 1680 cm-1
• CC 2200 cm-1

• Conjugation lowers the frequency:

• isolated C=C 1640-1680 cm-1
• conjugated C=C 1620-1640 cm-1
• aromatic C=C approx. 1600 cm-1

• Bonds with more s character absorb at a higher frequency.

Ranjana Prakash
Interpretation of IR spectra
SCBC-TIET

O-H and N-H Bonds

• Alcohol O-H
• broad with rounded tip when hydrogen bonding is present (sharp in
the absence of hydrogen bonding)

• Secondary amine (R2NH)

• Broad (usually) with one sharp spike

• Primary amine (RNH2)

• Broad (usually) with two sharp spikes.

Ranjana Prakash
Interpretation of IR spectra SCBC-TIET

Carbonyls
• Carbonyl stretches are generally strong:
• Aldehyde ~ 1710 cm-1
• Ketone ~ 1710 cm-1
• Carboxylic acid ~ 1710 cm-1
• Ester ~ 1730 - 1740 cm-1
• Amide ~ 1640-1680 cm-1

• Conjugation shifts all carbonyls to lower frequencies. 1745 cm

-1

• Ring strain shifts carbonyls to higher frequencies. O

H3C

Ranjana Prakash
 Interpretation of IR spectra SCBC-TIET

Infrared Absorption Frequencies

Functional Group Range (cm-1) Assignment
Alkanes 2850-3000 CH3, CH2 & CH
2 or 3 bands
Alkenes 3020-3100 =C-H & =CH2
1630-1680 C=C
1900-2000 C=C asymmetric stretch
Alkynes 3300 C-H
2100-2250 C≡C

Source: https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/InfraRed/infrared.htm
Ranjana Prakash
Interpretation of IR spectra
Infrared Absorption Frequencies SCBC-TIET

Functional Group Range (cm-1) Assignment

Arenes 3030 C-H
1600 & 1500 C=C (in ring) (2 bands) (3 if conjugated)
Alcohols & 3580-3650 O-H (free), usually sharp
Phenols 3200-3550 O-H (H-bonded), usually broad
970-1250 C-O
Amines 3400-3500 N-H (1°-amines), 2 bands
3300-3400 N-H (2°-amines)
1000-1250 C-N
Aldehydes & 2690-2840(2 bands) C-H (aldehyde C-H)
Ketones 1720-1740 C=O (saturated aldehyde)
1710-1720 C=O (saturated ketone)

1690 aryl ketone

1675 α, β-unsaturation
1745 cyclopentanone
1780 cyclobutanone
Ranjana Prakash
Interpretation of IR spectra
Infrared Absorption Frequencies SCBC-TIET

Functional Group Range (cm-1) Assignment

Carboxylic Acids & Derivatives 2500-3300 (acids) overlap C-H O-H (very broad)
1705-1720 (acids) C=O (H-bonded)
1210-1320 (acids) O-C

1785-1815 (acyl halides) C=O

1750 & 1820 (anhydrides) C=O (2-bands)
1040-1100 O-C
1735-1750 (esters) C=O
1000-1300 O-C (2-bands)
1630-1695(amides) C=O (amide one band)
Nitriles 2240-2260 C≡N (sharp)

Isocyanates,Isothiocyanates, 2100-2270 -N=C=O, -N=C=S

Diimides, Azides & Ketenes -N=C=N-, -N3, C=C=O

Source: https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/InfraRed/infrared.htm
Ranjana Prakash
 SCBC-TIET

In the next session…..

• Applications of IR spectroscopy

Ranjana Prakash