Spectroscopy

Powerful tool to analyse the atomic

and molecular structure
 Spectroscopy is the branch of science dealing with the
study of interaction of electromagnetic radiation with
matter. There are two main classes of spectra, absorption
spectra & emission spectra.
When white light is passed through prism / grating , it gets split
up into seven different colours. The set of colour thus obtained is
called as spectrum.
When white light is passed through the absorbing
substance and then observed through spectroscope,
some of the wavelengths were missing and dark lines
appear on those places. The resulting spectrum is
called absorption spectrum.
On the other hand, the light emitted by a substance is
passed through the prism and observed directly
through the spectroscope, the resulting spectrum is a
2 emission spectrum.
 Electromagnetic radiation is produced due to the interaction between the
electric and the magnetic field.

 Electromagnetic radiations are divided into various regions according to

their wavelength.

 The arrangement of electromagnetic radiation in the increasing order of

wavelength or decreasing order of frequency is known as Electromagnetic
spectrum.

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 There are two types of spectroscopy,
Atomic spectroscopy
Molecular spectroscopy
Atomic spectroscopy : It deals with
interaction of electromagnetic
radiation with atoms.
Atomic spectra It is a line spectra and occurs by the
electronic transition in an element.
 MOLECULAR SPECTRA:
It deals with the interaction of
electromagnetic radiation with molecules.
 It is a complicated spectra.

 It is due to the vibrational, rotational

and electronic transition in the
molecule.
Molecular spectra
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UV
SP -V
EC IS
TR IBL
OS E
CO
P
 Absorption radiation in the UV (200-400 nm) and visible (400-750 nm) regions of the
electromagnetic spectrum results in the excitation of valence electrons

Near UV region– 400 to 200 nm

Far UV region – 200- 10 nm

The electrons that are excited by the UV- Visible source are,

Electrons Compound Energy Type of Bond

Required

σ -- electrons Saturated long Less than 200 Single bond

chain & cyclic nm
hydrocarbons
π -- electrons Unsaturated UV / Visible Double / Triple
hydrocarbons bond

n -- electrons Compounds UV Unshared /

with hetero lone pair
atoms electrons
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 Generally an organic molecule consists of σ, π and non bonding
electrons (lone pairs). For example., the electrons of
formaldehyde is given below.

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 Terminologies in UV-Visible Spectroscopy

Chromophore

“ The presence of one or more unsaturated linkages

(π- electrons) in a compound is responsible for the
colour of the compound, these linkages are refered
to as chromophores.
They undergo π→π* transitions in the short
wavelength regions of UV-Visible spectra.

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 Non-conjugated alkenes show an intense absorption below 200 nm, and non
conjugated carbonyl compound shows a weak band around 200 - 300 nm.

1,5 – hexadiene has Acetone has

Ethylene has
λmax = 178 nm λmax = 279 nm
λmax = 171 nm

When double bonds are conjugated, λmax is shifted to longer

wavelength.
2,4 hexadiene has
λmax = 227 nm

Crotonaldehyde has
λmax = 290 nm

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 Auxochrome Benzene Has λmax = 255
nm

“
⊷ An atom or group of atoms which does
not gives rise to absorption band on its
own, but when conjugated to
chromophore it will cause a red shift.
⊷ Eg. –OH, NH2,-Cl,-Br,-I etc…

Phenol Has λmax = 270 nm

Aniline Has λmax = 280 nm

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 Shift in Wavelength &
Intensity
 Bathochromic shift – shift to
a longer wavelength
 Hypsochromic shift – Shift to
a shorter wavelength.
 Hyperchromic effect-
Increase in peak intensity
 Hypochromic effect –
Decrease in peak intensity

,
red
i e s are
r
y c her s are
W h be r r i e
e
Blu !!!!!!
e
bl u

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 This higher energy source (UV/Visible), 100- 100,000 KJ / mol simultaneously
causes rotational and vibrational transitions also. The energy change due to
electronic transition is greater than that of vibrational and rotational transitions,

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 Graphical representation of electronic transition in an organic molecule,

Where,σ  σ* & π  π* are bonding to anti bonding transitions.

n  σ* & n  π* are non-bonding to anti bonding
14 transitions.
 The energy requirements of the electronic transitions follows the order,
n→π*< π→π*< n→σ*< σ→σ*
n→π* transition requires lowest energy while σ → σ* requires highest
amount of energy.

 It is not essential that, the molecule on exposure to

UV / Visible radiation must give all kinds of
electronic transitions.

 The probability and intensity of the electronic

transition depends on the molar extinction co-
efficient (€) & other factors like effects of solvent ,
pH, conjugation etc..

 Based on the symmetry considerations, some of the

transition are allowed & some are forbidden.

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 σ  σ* Transitions Absorption bands are seen in the
far UV region. (Requires high
energy).
This region is less informative.

Occurs in saturated compounds

like methane, ethane etc..

The absorption is done under

vacuum condition, since O2 & N2
in the atmosphere will absorb @
far UV

λ max of CH4 for the σ  σ* is 121.9

nm. λ max of CH3-CH3 for the σ  σ*
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is 135 nm
 n → σ* Transition

Saturated Example,
compounds Trimethyl
It involves amine
with lone pair Absorption
promotion of (CH3)3N
of electrons bands appear
electron from shows two
undergo at longer
non-bonding absorption
n→σ* wavelength in
orbital (n) to bands.
transition in the near UV
antibonding
addition to region. n→σ* -
σ* orbital.
σ→ σ* 227nm
transition
σ→σ* - 99
nm

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 Aldehyde / ketone without double bond
shows n→π* transitions in the range
270-300nm
n→π* Transition
Here the unshared pair of electron /
lone pair is excited to antibonding π*
orbital . It is common in unsaturated
compounds with hetero atoms.

Aldehyde / ketone with It requires least amount of energy than all

unsaturation / double bond other transitions, hence the absorption band
shows n→π* transitions in
occurs at longer wavelength.
the range 300-350nm

As per symmetry considerations, it is a

forbidden transition, hence it is less intense.

Example, 3-Butene - 2 one, shows two

absorption band,
Low intensity band @ 324 nm - n→π*
High intensity band @ 219 nm - π→π*
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 π→π* TRANSITION

• These transitions are common

in unsaturated compounds like
alkene, alkyne, aromatics and
carbonyl compounds.

• Selection rule determines

whether transition to a
particular π*orbital is allowed
or forbidden.

• Example, λ max of ethylene for

π→π* transition is 171 nm.
λ max of 1,3 butadiene for π→π*
transition is 217 nm.

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 Most absorption bands of organic compounds are
due to electronic transitions from levels n or π to
the excited levels π* .
n→π* transitions have low molar
absorptivity (Є ̴ 10-100 L/mol.cm)
π→π* transitions have higher molar
absorptivity (Є ̴ 1000-10000 L/mol.cm .
Solvent often influences the position of
absorption bands, With the increase in polarity
n→π* transitions are shifted to lower wavelength
(Blue shift) and π→π* transitions are shifted to
longer wavelength (Red shift)

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 Chromophore
These are responsible for the It does not impart colour, but
colour of the compound.
It contains at least one
unsaturated bond. (-N=N-,
alkene, alkyne, ..)
Pale yellow coloured nitro
benzene becomes dark
yellow coloured when
hydroxyl group is attached
to it
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Auxochrome
when conjugate to
chromophore, it intensifies
the colour.
It contains lone pair (-OH,
NH2..)
Colourless benzene gets a
pale yellow colour, when
nitro group is attached to it.
 INSTRUMENTATION

Cells

Monochromator Detector

Source Recorder
Components

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 Block Diagram

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 Radiation Source

Tungsten / Hydrogen / Deuterium lamps are commonly

used as the radiation source.
 It must be stable & able to supply continuous
radiation.
 It must be with sufficient intensity.
Monochromator.

It is used to separate the radiations according to their wavelength

with the help of dispersing element & slits.
 Prism / grating is generally used as dispersing element.

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 Sample / reference cell
(Cuvette)

It is used to hold the sample and reference material.

The cell must be constructed of with the material, that is transparent to desired
electromagnetic radiation and inert towards the solvent used.
For visible spectra, cell made of glass/plastic are generally suitable.
For Ultraviolet region, quartz cells are preferred

Beam splitter

In case of single beam spectrophotometer, only one light beam /

optical path passes through the sample and reaches the detector.

In case of double beam spectrophotometer, light from the source

split into two beams with the help of beam splitter, one for the
sample and the other for reference.

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 Detectors

The detector senses the intensity of transmitted light & convert light
radiations into electrical signals.
 Barrier layer cell, photomultiplier tube & photodetectors are the
commonly used detectors.
Recorder

Recorder receives the signal from the detector amplifies it & gives the
result in the form of spectrum.

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 Working
 Light radiation from the source is irradiated through the
monochromator.
 The monochromator helps in passing a narrow beam of light
with the dispersing element.
 A narrow beam of light from the monochromator is split into
two halves by the beam splitter.
 From the beam splitter, one half of the beam passes through
the sample and the other through the reference cell.
 If the sample absorbs light at a particular wavelength, then the
intensity of the sample beam will be less than the reference.
 In case of single beam spectrophotometer, decrease in light
intensity with initial light (I o) is noted.
 The detector detects the intensity of light absorbed /
transmitted and converts the light signal into electrical signal.
The resulting electrical signal is amplified by amplifier.
 Finally an absorption spectra ( a plot of wavelength Vs
absorbance ) is shown in the monitor with the help of software
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28
 Identification
of conjugated
electrons

Studying Detection of
chemical functional
kinetics groups

Applications

Checking Distinguishing
purity & geometrical
Detection of isomers &
metal ions tautomers

Qualitative &
Quantitative
analysis

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 IR SPECTROSCOPY

 Infrared spectroscopy is one of the important

analytical techniques of scientist
today.
 It is a absorption spectroscopy which deals
with the interaction of Infrared region
of the electromagnetic spectrum with the
molecule.
 Infra red ranges from 12500 to 50 Cm -1 in
the electromagnetic spectrum.

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 Types of Molecular Vibrations

Absorption of Infrared radiation causes vibrational changes in a

molecule. Hence it is also called vibrational spectroscopy. The
fundamental vibrations are given below,

During stretching
bond length During bending
changes, but angle vibrations atoms move
remains unaltered in & out of the bond
axis- Bond length
remains same, bond
angle changes.

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32
 The vibrational frequencies can be calculated as it relates to bond
strength and masses of atoms attached that undergo vibration (Hooke’s
law)
K = force constant,
bond order or bond
strength
µ is reduced mass

From the above equation it is apparent that stronger the

bond higher the vibrational frequency.
Higher the masses of atoms lower the vibrational
frequency.

Energy of stretching is related to

hybridisation. Sp > sp2 > sp3

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 E streching > E bending

Selection Rules

For a particular vibration to be infrared active there must be a

change in the dipole moment of the molecule during the
vibration.
Thus homonuclear diatomic molecules like H 2, O 2, N 2 are
inactive in the infrared spectrum. & Heteronuclear diatomic
molecule such as CO, NO are active in IR.
In case of symmetrical polyatomic molecules such as CO 2, the
symmetric stretching vibration is infrared inactive where as the
34 asymmetric stretching vibration is IR active.
 ∆ν = ± 1, transition can take place between Adjacent vibrational
levels, 0 to 1, 1 to 2 etc.
IR spectrum shows bands rather than line spectrum due to
coupling of various rotational transitions within a given
vibrational transition .
IR spectrum is generally complex and contains many bands in
addition to the ones corresponding to fundamental vibrational
transitions.
Overtones: Bands corresponding to integral multiple of
fundamental vibration. They are due to transition from ground
state to higherCalibration
vibrationalof states.
IR
spectrum
A thin film of
polystyrene is used as a
standard for calibration
Peaks at 1603 and 3062
35 cm-1 are used to
 No. of Vibrational degrees of freedom

 For a non-linear molecule containing n atoms - the number of vibrational

degrees of freedom is (3n-6). Eg. H2O, CH4, C6H6
 For example, water molecule has ( 3 x 3 -6 = 3) three vibrational degrees of
freedom corresponding to two stretching mode and one bending mode of
vibration

 For a linear molecule containing n atoms the number of

vibrational degrees of freedom is (3n-5). Example HCl, HBr,
CO2, CO, NO..
 For example CO2 molecule has( 3x3-5 =4) four degrees of
vibrational freedom corresponding to two Stretching mode and
two bending modes of vibration.

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 The main use of IR spectroscopy is structural analysis, which is interpreted using
FUNCTIONAL GROUP REGION (4000 – 1500 Cm-1) :
Peaks in this region are characteristic of specific kinds of bonds, and therefore can be used to
identify the presence of specific functional group .

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 FINGER PRINT REGION (1500 – 400 Cm-1)
 In IR, the region below 1500 Cm-1 is rich in many absorption
bands
and it is called as finger print region.
 Peak in this region arises by the small change in structure or
deformation.
 They may be characteristic of molecular symmetry or
combination
bands arising from the deformation of multiple bonds
INTERPRETATION
simultaneously.  The normal way to interpret IR spectra is to
examine the the functional group region to
determine which groups might be present, then to
note any unusually strong bands or particularly
prominent patterns in the fingerprint region.
 Finally, if you think you have identified the
compound, you can compare the spectrum with a
reference.
38  Matching the fingerprint region is a very rigorous
 INSTRUMENTATION The components of IR Spectrometer are,1) Radiation source 2)
Monochromator 3) Cells 4) Detector 5) Recorder .
Radiation Source : Nernst glower : It consists of a rod of the sintered mixture of oxides
of Zr, Y, Er. This rod is electrically heated to 1500oC to produce IR radiation
Nichrome Wire : It is a non magnetic alloy of Ni & Cr
Monochromator : It allows the light of required wavelength to pass through.
Sample cells : It must be transparent to IR radiation , usually pellet made of KBr, NaCl
is used.
Detector : It measures the intensity of transmitted radiation. Common detectors-
Thermocouple, Pyroelectric detectors, photoconductivity cell.
Recorder : It records the resulting signal in the form of IR spectra

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40
 Identification
of functional
group

Research & Checking

Environmental purity &
application kinetics

Differentiating Applications Hydrogen

Geometrical
bond
isomers &
determination
Tautomers

Determination Qualitative &

of molecular Quantitative
symmetry analysis

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 ATOMIC ABSORPTION
SPECTROSCOPY

Analytical technique – Qualitative

& Quantitative analysis of trace
 s
h oi ?
W ?? ?
?
he

Alan
Walsh

Invented AAS in
1950’s
 The superiority of this technique is, it can analyse 50 – 60 elements on a
single run without any interference.
PRINCIPLE :
 AAS is based on atomisation of the sample followed by absorption
of characteristic radiation by the ground state gaseous atoms.
 When a solution having a mixture of metallic species is introduced
into the flame, the solvent evaporates and vapor of metal atoms
were obtained.
 when a light of particular
wavelength is passed through the
flame containing atoms of the
metallic species, part of the light
will be absorbed, which is
α
n
i o ti o
n measured using detector
t
p a
sor nt r  Thus in Atomic absorption
e
Ab nc spectroscopy, amount of light
Co
absorbed is determined
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 INSTRUMENTATION
Radiation Source: (Hallow Cathode lamp)
The radiation source should emit stable & intense characteristic radiation of the element
to be determined.
The hallow cathode lamp – a glass tube containing argon (anode) & hallow
cathode (made of analyte metal) is generally used as source.

Chopper : A rotating wheel is placed between hallow cathode lamp & flame. It
breaks the steady light from the lamp into an pulsating light . ( since recorder
record only the pulsating current (AC))

Burner / Flame : The flame is used for converting the liquid

sample into gaseous state (atomic vapor).

Nebulisation: Before the liquid sample enters the burner, it is converted into
small droplets by a process called nebulisation.

Monochromators : It selects the light of suitable wavelength from the hallow cathode
lamp. Eg – Prisms, Gratings

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Detector : Photomultiplier tube is the commonly
used detector, when the photon strikes it, an emf
is produced.

Read-out : The amplified signal from the

amplifier is recorded in the monitor.
 Working :
 The characteristic radiation from the
hallow cathode lamp is passed through
the flame into which the sample is
aspirated.
 The metallic compounds are decomposed
Place your screenshot here
into atoms of the element to be measured.
 The atoms absorbs characteristic radiation
in the flame.
 The unabsorbed radiation from the flame
is passed through the monochromator
followed by its detection in the detector.
 Finally the data from the detector is
Applications recorded in the form of spectra.
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HAPPY
LEARNING
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