Spectroscopy

Introduction

It is the study of interaction

between matter and radiated energy.
Light interacting with matter as an
analytical tool.
 Different Spectroscopies

• UV-visible :– electronic states

of valence e/d-orbital
transitions for solvated
transition metals
• Fluorescence :– emission of
UV/visible by certain
molecules
• FT-IR :– vibrational transitions
of molecules
• FT-NMR :– nuclear spin
transitions
 Types of material

• Spectroscopic
studies are
designed so
that the
radiated
energy
interacts
with
specific
types of
 Atoms

• Atomic absorption
spectroscopy (AAS) involve
visible and ultraviolet
spectroscopies.
• Atoms of different elements
have distinct spectra and
therefore atomic
spectroscopy allows for the
identification and
 Molecules
• The combination of
atoms into
molecules leads
to the creation
of unique types
of energetic
states and
therefore unique
spectra of the
transitions
between these
states. Molecular
 Nuclei
• Nuclei also have
distinct energy
states that are
widely
separated and
lead to gamma
ray spectra.
Distinct
nuclear spin
states can have
their energy
separated by a
Electronic Excitation by UV/Vis Spectroscopy :
UV:
valanc IR: Radio waves:
X-ray:
core e moleculNuclear spin states
electron electr ar (in a magnetic fiel
excitati onic vibrati
on excita ons
tion
  Spectroscopic Techniques and
 

Chemistry they Probe

UV-vis UV-vis Bonding  

Atomic Absorption region

UV-vis electrons
Atomic
FT-IR region
IR/Microw transitions
Vibrations,
Raman ave
IR/UV rotations
Vibrations  
FT-NMR Radio Nuclear spin
X-Ray Spectroscopy waves
X-rays states
Inner electrons,
X-ray Crystallography X-rays elemental
3-D  
structure
  Spectroscopic Techniques and Common Uses
 

UV-vis UV-vis Quantitative  

Atomic Absorption region
UV-vis analysis /Beer’s
Quantitative
region Law
FT-IR IR/Microw analysis
Functional Group
ave Beer ’s Law
Analysis
Raman IR/UV Functional Group 
Analysis/quant
FT-NMR Radio Structure
X-Ray Spectroscopy waves
X-rays determination
Elemental
X-ray Crystallography X-rays Analysis
3-D structure  
Anaylysis
UV and Visible Spectroscopy

• absorption spectroscopy
or
reflectance
spectroscop
y in the
ultraviolet
-visible
spectral
region
The Spectroscopic Process
1.In UV spectroscopy, the sample is
irradiated with the broad spectrum
of the UV radiation
2.If a particular electronic transition
matches the energy of a certain band
of UV, it will be absorbed
3.The remaining UV light passes through
the sample and is observed
4.From this residual radiation a
spectrum is obtained
π∗
with “gaps” at
these discrete energies – this is
called an absorption spectrum
5.
π∗
6. π

π
13
The UV Absorption
process
•  * and   * transitions: high-
energy, accessible in vacuum UV
( max <150 nm). Not usually observed
in molecular UV-Vis.
•n  * and   * transitions:
non-bonding electrons (lone pairs),
wavelength (max ) in the 150-250 nm
region.
•n  * and   * transitions:
most common transitions observed in
organic molecular UV-Vis, observed in
compounds with lone pairs and
multiple bonds with max = 200-600
nm.
•Any of these require that incoming
 π* π* π*
Nature of π* π* π*
Examp
le
π* π* π*
these n n n
for a
simpl
absorptions π π π
π
e
enone
π π
-*; max =218 n- *;
=11,000  max =320
=100
Example:   * transitions responsible for
ethylene UV absorption at ~170 nm calculated with
ZINDO semi-empirical excited-states methods
(Gaussian 03Ω ):
h 170nm photon

LUMO g antibonding molecular orbital
HOMO u bonding molecular orbital
 The Quantitative Picture
•Transmittance :
P0 P
T = P/P0 (power in) (power out)

•Absorbance :
A = -log10 T = log10 P0/P B(path through sample)

•The Beer-Lambert Law (a.k.a. Beer’s Law):

A = ebc
Where the absorbance A has no units, since A =
log10 P0 / P
e is the molar absorbtivity with units of L mol-1
cm-1
b is the path length of the sample in cm
 slit cuvette
source
detector
BEER’S LAW
“The
•

absorption of
l;ight by a
substance of
particular
wavelength is
proportional
to the number
of molecules
in the path
of light.”
•

•
 Chromophores and Auxochromes
Chromophres :
ated groups responsible for - * and n→ * electroni
ons. e.g. C=C , C=O , N=N and N=O ( 200nm-800nm)
-Auxochromes
Are saturated groups posses unshared
electrons, and does
3)Bathochromic Hypsochromic
not absorb in&near shiftradiations
UV or visible
e.g. OH,NH2.
-Bathochromic shift (or red hift)
It is the shift of max to a longer wavelength
due to substitution with certain functional
groups (e.g. –OH and –NH2), when two or more
chromophores
sochromic shift are present
(or blue in) conjugation, change
shift
in pH and effect of the medium (solvent).
It is the shift of max to a shorter
wavelength due to removal
yperchromic effect
an increase in the intensity of
absorption usually due to
introductioneffect
Hypochromic of an auxochrome
ves a decrease in the intensity of absorption
-Auxochromes
when attached to chomophoric
molecule, increase both its
wave length and intensity of
absorption
Because maximum inters
auxochrome . into resonance
interaction with the chromophore , thus
increase the extent of conjugation, shift
the absorption maximum to longer wave
Effect of pH
The spectra of compounds containing acidic (phenolic-OH)
or
basic (-NH2) groups are dependent on the pH of the
medium. The U.V spectrum of phenol in
acid medium, benzenoid form
while in alkaline medium is the
phenate anion ,quinonoid form

The free pair of of electrons

of O2 increasing the
elocalization of the  -
electrons, leading to the
formation of conjugated system.
So , electrons become more
energetic and need less energy
to be excited, therefore absorb
Phenol longer  bathochromic shift ;
red shift) with hyperchromic
effect
 Applications
• UV/Vis spectroscopy is routinely
used in analytical chemistry for
the quantitative determination of
different analytes, such as
transition metal ions, highly
conjugated organic compounds, and
biological macromolecules.
• UV/Vis spectroscopy can be used to
determine the concentration of
the absorber in a solution.
• The wavelengths of absorption peaks
can be correlated with the types
of bonds in a given molecule and
 Infrared Spectroscopy
• Infrared
spectroscopy
is the
spectroscopy
that deals
with the
infrared
region of the
electromagnetic spectrum
, that is
light with a
longer
 IR-REGION: 12,800 - 10 cm -1

REGION WAVE WAVE FREQUENCY

LENGTH NUMBER RANGE
λ υ ( cm -
( μm ) 1) Hz
NEAR 0 . 78 12800 - 3 . 8x10 14 -
- 2.5 4000 1 . 2x10 14
MIDDLE 2.5 - 4000 - 200 1 . 2x10 14
50 - 6x1 12
FAR 50 - 200 - 10 6x10 12 -
1000 30x10 11
MOST 2 . 5 - 15 4000 - 670
USED 1 . 2x10 14 -
2x10 13
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•BASIC PRINCIPLE
•

• IR involves absorption phenomenon the

absorption of radiation depends on
increasing energy of vibration or rotation
associated with covalent bond in molecule
provide that such an increasing in energy
causes a change in the dipole moment of
molecule.
• Hence in order to absorb IR radiation a
molecule must go a net change in dipole
moment due to its vibration or rotation
motion.
• This means that nearly all molecules
containing covalent bond will show some
degree of selective IR absorption .
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n
nThis makes absorption band appear as dips in
the curve than as maxima as in the case of
UV-Vis.
nEach dip In the spectrum is called a band or
peak and represented absorption of IR
radiation at that frequency by the sample.
nThe transmittance is 0% if all the
radiation is absorbed and the transmittance
is 100% for no absorption.

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 Radiation sources

1.Nernest glower
2.Globar source
3.Incandescent wire source
4.Mercury arc
5.Carbon dioxide laser source
6.Tungsten filament lamp

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•Use of Wave number than
wavelength offer several
advantages
nWavenumber are directly proportional
to frequency and are expressed in
much more convenient numbers (in
this region of the spectrum), 5000-
500 cm-1

nBecause the wave number is directly

proportional to frequency and energy,
the use of wave numbers allows
spectra to be displayed linear in
energy, which is a distinct aid in
sorting out related vibrational
transitions
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 INFRARED THEORY
- Molecular vibration can occur by 2
different mechanism
- Firstly, quanta of IR radiation can
excite
atoms to vibrate directly. The
absorption of
IR radiation give rise to the IR
spectrum
- Secondly quanta of visible light
achieve the
same result indirectly
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 Types
of
Vibrati
Stretch on
ing Bending

Sciss
Symmet oring
Wagg
ric In plane ing

Rocki Out of plan

Asymm ng Twist
etric ing31
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 Molecular Vibrations

• Covalent bonds vibrate at only

certain allowable frequencies.

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 Stretching Frequencies

• Frequency decreases with increasing

atomic weight.
• Frequency increases with increasing
bond energy.
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 Vibrational Modes

• Nonlinear molecule with n atoms

usually has 3n - 6 fundamental
vibrational modes.

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 Fingerprint of Molecule
• Whole-molecule vibrations and bending
vibrations are also quantitized.
• No two molecules will give exactly
the same IR spectrum (except
enantiomers).
• Simple stretching: 1600-3500 cm-1 .
• Complex vibrations: 600-1400 cm-1 ,
called the “fingerprint region.”

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 IR-Active and Inactive
• A polar bond is usually IR-active.
• A nonpolar bond in a symmetrical
molecule will absorb weakly or
not at all.

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 n-pentane

2850-2960 cm-1
3000 cm-1 sat’d C-H

1470 &1375 cm-1

CH3CH2CH2CH2CH3
n-hexane

CH3CH2CH2CH2CH2CH3
cyclohexane

no 1375 cm-1
no –CH3
 Applications
• Identification of all types of organic &
many types of inorganic compounds
• Determination of functional groups
• Identification of chromatographic
effluents
• Quantitative determination of compounds
in mixtures
• Determination of molecular composition &
stereochemistry
• Determination of molecular orientation
(Polymers & Solutions)
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• Identification of compounds by matching
unknown with reference
spectrum(Fingerprinting)
• Detection of impurities 0.1-0.01
• Analysis of formulations such as
insecticides & copolymers
•Accuracy

§ Favorable conditions >1%

§ Routine analysis ±5%

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 Sensitivity & Detection
limits
• Routine – 2%
• Most favorable conditions & using
special techniques – 0.01%
•Limitations

§ Minimal element info.is given for most

of samples
§ Background solvent should be
transparent in
• IR region
§ Molecule must be active in IR region
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