SPECTROSCOPY
It is the branch of science that deals with the study of interaction of
matter with light.
OR
It is the branch of science that deals with the study of interaction
of electromagnetic radiation with matter.
Electromagnetic Radiation
1. Electromagnetic radiation consist of discrete packages of
energy which are called as photons.
2. A photon consists of an oscillating electric field (E) & an
oscillating magnetic field (M) which are perpendicular to each
other.
Interaction of EMR with matter
1. EMR interacts with electronic energy levels or vibrational
energy levels or rotational energy levels.
2. The fundamental process of absorption of energy by a
molecule is based on the absorption of a discrete amount of
energy.
3. The energy required for the transition from a lower state to
higher state is equal to the energy of EMR
The order of energy absorbed is;
∆E
electronic >∆Evibration > ∆Erotation
PRINCIPLE OF ABSORPTION SPECTROSCOPY
It is given by Lambert-Beer’s Law.
Statement:
When a monochromatic light is passed through a dilute solution of a
sample, the decrease in the intensity of radiation with the thickness
of the solution is directly proportional to the intensity of transmitted
light as well as the molar concentration of the solution.
Lambert-Beer’s Law:
Where
I0 = intensity of incident light
I= intensity of transmitted light
Mathematically, it can be expressed as A = ε c l
Where,
A = absorbance or optical density
ε = molar absorption coefficient or absorptivity
c = molar concentration of sample
(mol/L or mol/dm3)
l = thickness of the solution in cm
Derivation
According to the statement,
dI /dx α I . c…………. (1)
or
dI / I = kc dx………….. (2)
dI /dx= change in intensity w.r.t thickness
On integration within the limit we get,[(I0 to I) and 0 to x]
-ln I/I0= = kcl…………………….(3)
or
ln I0/ I = kcl……………. (4)
or
2.303 log I0/ I = kcl……………..(5)
log I0/ I = (k/ 2.303 ) cl……….. (6)
log I0/ I A = absorbance and
(k/ 2.303 ) =ε(molar absorption coefficient)
Then equn. 6 becomes, A = ε c l
Again, transmittance(T) =I / I0…………………. (7)
then,-log T = ε c l
T = 10 - ε c l= 10 – A……………………… (8)
Limitation of the law :
1. It is valid for very dilute solution.
2. Appropriate solvent must be used.( highly pure non polar
solvents do not interact with sample molecules where as polar
solvents may form hydrogen bond.)
3. The temp. should not vary to a large extent.
4. This is valid for maximum absorbance of 1.
NOTE
1. Absorbance or optical density, A = log I0 / I ( unit less)
2. Transmittance(T) = I / I0 = 10- ε c l = 10 -A ( unit less)
3. Molar absorption coefficient or absorptivity, ε = A /cl
(mol-1dm3cm-1 o)
4. Molar concentration ,= c (mol/L OR mol/dm3)
5.Thickness or path length or cell length,= l (cm)
Example-1
A light of wave length 500nm is allowed to pass through a
cell of 2 mm path Length containing 10-3mole/dm3 of
compound X.If the absorbance of this Solution is 0.5
then .calculate the molar absorption coefficient and
transmittance
Ans.
Given A=0.5
C= 10-3 mole/dm3
l=0.2cm
A=Absorption coefficient =εcl
Absorption coefficient (ε) =A/cl
=0.5/ 10-3 mole/dm3 x 0.2cm=2.5x103
A=logIo/I=0.5
T=I/Io=10-0.5=1/100.5=0.3162
Basic Principle of UV-VISIBLE Spectroscopy
1. Absorption of UV-VISIBLE raditation by a molecule results in
electronic transition from occupied molecular
orbitals(bonding molecular orbitals) of lower energy to
unoccupied molecular orbitals(antibonding molecular
orbital) of higher energy.
2. σ, π and n(bonding molecular orbitals) to σ* , π* (antibonding
molecular orbitals)
3. The common solvent used for preparing sample to be
analyzed is either ethyl alcohol or hexane.
Types of Electronic Transitions
It involves the transition of electrons from one of the
3-ground states (σ, π and n) molecular orbitals to one
of the two excited states(σ* , π* ) antibonding molecular
orbitals.
There are 4-types of transitions possible among the
molecular orbitals
1- σ → σ* Transitions
2- n€→ σ* Transitions
3- π → π* Transitions
4- n€→ π* Transitions
1.
σ → σ* Transitions(<165nm)
1. σ electron from bonding orbital is excited to
corresponding anti-bonding orbital σ*.
2. The energy required is large for this transition.
e.g. Methane (CH4) has C-H bond only and can undergo σ →
σ* transition and shows absorbance maxima at 125 nm.
2.
π → π* Transitions(>165nm)
1. π electron in a bonding orbital is excited to corresponding
anti-bonding orbital π*.
2. Compounds containing multiple bonds like alkenes, alkynes,
carbonyl, nitriles, aromatic compounds, etc undergo π → π*
transitions.
e.g. Alkenes generally absorb in the region 170 to 205 nm.
3.
n€→ σ* Transitions(>185nm)
1. An electron from non-bonding orbital is promoted to
anti-bonding σ* orbital.
2. Saturated compounds containing atoms with lone pair
of electrons like O, N, S and halogens are capable of n
→ σ* transition.
3. These transitions usually requires less energy than σ →
σ* transitions.
n€→ π* Transitions (>270nm)
1. An electron from non-bonding orbital is promoted to
anti-bonding π* orbital.
2. Compounds containing double bond involving hetero
atoms (C=O, C≡N, N=O) undergo such transitions.
3. n → π* transitions require minimum energy and show
absorption at longer wavelength around 300 nm .
Conclusion
Thus, n → π* & π → π* electronic transitions show
absorption in region above 200 nm which are accessible to
UV-visible spectroscopy.
In case of n → π* transition system
The absorption maxima is shifted to shorter wave length ( blue shift)
in polar solvent due to the formation of hydrogen bond with the
ground state of the molecule .
Acetone contains n → π* transition
Ex-1 The absorption maxima of acetone in hexane is 279 nm
but in water it is shifted to 264 nm.
Hexane-nonpolar solvent
Water-polar solvent
In case of π → π* transition system
The absorption maxima is shifted to longer wave length ( red shift)
in polar solvent because the excited states are more polar than
the ground state
Alkenes contain π → π* transition
Ex-2. Alkene shows red shift in polar solvent (λmaxis increased by
10-20 nm.)
Effect of π-conjugation
1. Increase in conjugation increases the λmax towards longer
wave length.
(Conjugation means alternative arrangement of π bond in
a series)
TERMS USED IN UV-VISIBLE SPECTROSCOPY
Chromophore:
The part of a molecule responsible for imparting color, is
called as chromophore.
OR
The functional groups containing multiple bonds capable of
absorbing radiations above 200 nm due to n → π* & π → π*
transitions.
e.g. NO2, N=O, C=O, C=N, C≡N, C=C, C=S, etc
Auxochrome
The substituent that does not absorb UV radiation but its
presence (when attached to a chromophore) modifies the
ability of the chromophore by altering the wave length and
intensity of absorption is called auxochrome.
e.g. methyl (- CH3), ethyl (-C2H5), hydroxyl (-OH), halogen
(-X), amino (-NH2), alkoxy (-OR) etc.
effect
Benzene λmax = 255 nm,absent of auxochromic group
Phenol λmax = 270 nm,OH is auxochromic group
Phenol has higher λmax value due to the presence of OH
auxochromic group
Hypsochromic shift;
It is an effect due to which the absorption maximum is
shifted towards shorter wave length (blue shift)and it may
be due to the removal of conjugation or change in polrity
of solvent.
Hyperchromic shift or effect;
When absorption intensity (ε) of a compound is increased, it is
known as hyperchromic shift. If auxochrome introduces to the
compound, the intensity of absorption increases.
Exam-1
Pyridine
λmax = 257nm
ε = 2750 (intensity of absorption)
2-methyl pyridine
λmax = 260 nm
ε = 3560(intensity of absorption)
Hypochromic shift or effect;
When absorption intensity (ε) of a compound is decreased, it is
known as hypochromic shift.
APPLICATIONS OF UV -VISIBLE SPECTROSCOPY
1. It is used for characterizing aromatic compounds and
conjugated olefins.
2. It can be used to find out molar concentration of the
solute under study.
3. -Detection of impurities:It is one of the
important method to detect impurities in
organic solvents.
4. Detection of isomers are possible.
5. Determination of molecular weight using Beer’s law.( -
dI /dx α c)
