UV-VISIBLE SPECTROSCOPY

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 INTRODUCTION

Spectroscopy is a technique that measures the interaction of molecules with

electromagnetic radiation.
 Light in the near-ultraviolet (UV) and visible (vis) range of the electromagnetic spectrum
has an energy of about 150– 400 kJ mol.
The energy of the light is used to promote electrons from the ground state to an excited
state.
A spectrum is obtained when the absorption of light is measured as a function of its
frequency or wavelength.

The absorbance of a solute depends linearly on its concentration and therefore absorption
spectroscopy is ideally suited for quantitative measurements.

Spectroscopic measurements are very sensitive and nondestructive, and require only small
amounts of material for analysis

Application of derivative technique of spectrophotometry offers a powerful tool for

quantitative analysis of multi-component mixtures.

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 Principle and Basic concept
Ultraviolet – visible spectroscopy (λ 200 - 800 nm) studies the changes in electronic
energy levels within the molecule arising due to transfer of electrons from π- or non-
bonding orbital's.
It commonly provides the knowledge about π-electron systems, conjugated unsaturations,
aromatic compounds and conjugated non-bonding electron systems etc
UV- Visible is divided into the ultraviolet (UV, 190-400 nm) and visible (VIS, 400-800
nm) regions. Since the absorption of ultraviolet or visible radiation by a molecule leads
transition among electronic energy levels of the molecule, it is also often called as
electronic spectroscopy.

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 Nature of Electronic Transitions
The total energy of a molecule is the sum of its electronic, its vibrational energy and its
rotational energy.
Energy absorbed in the UV region produces changes in the electronic energy of the
molecule. As a molecule absorbs energy, an electron is promoted from an occupied
molecular orbital (usually a non-bonding n or bonding π orbital) to an unoccupied
molecular orbital (an anti-bonding π∗ or σorbital) of greater potential energy (figure1).

Fig.1)Relative energies of orbitals most commonly involved in electronic spectroscopy of organic

molecules
increasing order of their energies viz. n→π* < n→σ* < π →π* < σ→π* < σ →σ*
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Principles of Absorption Spectroscopy: Beer’s and Lambert’s Law
The greater the number of molecules that absorb light of a given wavelength, the greater the
extent of light absorption and higher the peak intensity in absorption spectrum .
This makes the basis of Beer-Lambert Law which states that the fraction of incident
radiation absorbed is proportional to the number of absorbing molecules in its path .
When the radiation passes through a solution, the amount of light absorbed or transmitted is
an exponential function of the molecular concentration of the solute and also a function of
length of the path of radiation through the sample. Therefore,
Log Io / I = ε c l
Where Io = Intensity of the incident light (or the light intensity passing through a reference
cell)
I = Intensity of light transmitted through the sample solution
c = concentration of the solute in mol l-1
l = path length of the sample in cm
ε = molar absorptivity or the molar extinction coefficient , ε is numerically equal to the
absorbance of a solution of unit molar concentration (c = 1) in a cell of unit length ( l = 1)
and its units are liters. moles-1. cm-1.
The ratio I / Io is known as transmittance T and the logarithm of the inverse ratio Io / I is
known as the absorbance A. Therefore
- Log I / Io = - log T = ε c l
and Log Io / I = A = ε c l
o6rA = ε c 19-May-19
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Spectral Measurements
The UV-Vis spectra are usually measured in very dilute solutions and the most
important criterion in the choice of solvent is that the solvent must be transparent
within the wavelength range being examined.
Table lists some common solvents with their lower wavelength cut off limits. Below
these limits, the solvents show excessive absorbance and should not be used to
determine UV spectrum of a sample.
SR.NO. Solvent Cut off
wavelength(nm)
1 Acetonitrile 190
2 Water 191
3 Cyclohexane 195
4 Methanol 203
5 95% ethanol 304
Of the solvents listed in table , water, 95% ethanol and hexane are the most commonly
used solvents.
For recording the spectrum 1 cm square quartz cell is commonly used. These require
approx. 3 ml of solution.
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Applications

Application to organic compound

 Detection of conjugation
 Detection of geometrical isomers
 Detection of functional group
 Qualitative analysis
 Detection of impurities
 Quantitative analysis
 Chemical kinetics

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Conclusions

UV-visible spectroscopy, a simple, rapid, precise and highly accurate method for
quantitative estimation is in great use now a day. Derivative spectrophotometry is an
analytical technique of great utility for extracting both qualitative and quantitative
information from spectra composed of unresolved bands by calculating and plotting one of
the mathematical derivatives of a spectral curve. Therefore the derivative spectra (first to
fourth-order) of the mixtures were checked to select a suitable spectrum to be used for the
simultaneous determination of the components.
Derivative techniques in spectroscopy often offer a powerful tool for a resolution
enhancement, when signal overlaps or interference exists. Several specific signals were
singled out for the components in the spectra of different derivative orders but the first-order
derivative spectra seemed to be generally the most suitable for analytical aim.
A derivative spectrum shows better resolution of overlapping bands than the fundamental
spectrum and may permit the accurate determination of the λmax of the individual bands.
Secondly, DS discriminates in favor of substances of narrow spectral bandwidth against
broad bandwidth substances. All the amplitudes in the derivative spectrum are proportional
to the concentration of the analyte provided that Beer's law is obeyed by the fundamental
spectrum.

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