The Absorption of Radiation

(UV and Visible)

▪ If a beam of white light passes through a glass container
(cuvet) filled with liquid, the emergent radiation is always less
powerful than that entering.
▪ The diminution of power is generally of different extent for
different wavelengths.
▪ The loss is due to in part to reflections in the surfaces and to
scattering by any suspended particles present, but at the
absence of such particle, it is primarily accounted for by the
absorption of radiant energy by the liquid.
▪ If the energy absorbed is greater for some visible wavelengths
than for others, the emergent beam will appear colored.
Components of total attenuation process
 Colors of Visible Radiation
Approximate wavelength range, nm Color Complement
400-465 Violet Yellow-green
465-482 Blue Yellow
482-487 Greenish blue Orange
487-493 Blue-green Red-orange
493-498 Bluish-green Red
498-530 Green Red-purple
530-559 Yellowish green Reddish purple
559-571 Yellow-green Purple
571-576 Greenish yellow Violet
576-580 Yellow Blue
580-587 Yellowish orange Blue
587-597 Orange Greenish blue
597-617 Reddish orange Blue-green
617-780 Red Blue-green
▪ The apparent color of the solution is always
the complement of the color absorbed.

▪ Thus a solution absorbing in the blue region

will appear yellow, one that absorps green
will appear purple, etc.
▪ The radiation absorbed by the solution is
characteristic of the material doing the
absorbing.
▪ A solution containing the hydrated copper
(II) ion absorbs yellow and is transparent
to blue, so copper may be determined by
measuring the degree of absorption of
yellow light under standardized
conditions.
▪ Any soluble colored material may be
determined quantitatively in this way.
▪ A substance that is colorless or only
faintly colored may often be determined
by adding areagent that will convert it to
an intensely colored compound.

▪ Addition of ammonia to a copper solution

produces a much more intense color that
that of the hydrated ion itself, and
therefore provides a more sensitive
analytical test.
▪ The chemical group most strongly influenching
molecular absorption characteristics is called a
chromophore.
▪ Chromophore which can be detected bu UV/Vis
spectrophotometers always involve a multiple
bond (such as C=C, C=O or C≡N) and may be
conjugated with other groups to form complex
chromophores.
▪ A typical example is the benzene ring which has
an absorption peak at 254 nm (λmax = 254 nm).
UV/Vis Spectrophotometer
General Optical Spectrometer
Single Beam
Double Beam
 Double Beam

The wavelength-dependent functions of the instrument are

significantly reduced to give much improved operating
characteristics by feedback system in the reference channel that
adjusts the detector gain to compensate for source and detector
variations.
Prism monochromator
Prism monochromator
Diffraction grating monochromator
Sample Cuvettes

The most commonly

used cuvett has a light
path length of 10 mm.
Longer or shorter path
lengths are useful if
concentration fall
outside normal ranges
without further
processing.
Types of Cuvett
 UV sources

 Arc discharge lamps with electrical discharge

maintained in appropriate gases.
 Low pressure hydrogen and deuterium lamps.
 Lasers - narrow spectral widths, very high
intensity, spatial beam, time resolution, problem
with range of wavelengths.
 Discrete spectroscopic- metal vapor & hollow
cathode lamps.
UV/Vis light source

UV
Vis
Absorption Peak (λmax)
 Sample Analysis

Percent transmittance plotted against concentration.

 Sample Analysis

Absorbance plotted against concentration.

Calibration Curve
Multicomponents System
 at λ1 : AIλ1 = εIλ1 bcI and AIIλ1 = εIIλ1 bcII
Aλ1 = AIλ1 + AIIλ1 = εIλ1 bcI + εIIλ1 bcII (1)

at λ2 : AIλ2 = εIλ2 bcI and AIIλ2 = εIIλ2 bcII

Aλ2 = AIλ2 + AIIλ2 = εIλ2 bcI + εIIλ2 bcII (2)
where:
Aλ1 and Aλ2 = absorbance of the mixture measured at λ1 and λ2
AIλ1 and AIλ2 = absorbance of component I in the mixture at λ1 and λ2
AIIλ1 and AIIλ2 = absorbance of component II in the mixture at λ1 and λ2
εIλ1, εIλ2, εIIλ1, and εIIλ2 = molar absortivity of components I and II at λ1
and λ2
cI and cII = concentration of components I and II in the mixture.
 n components

If there are n components in the mixture, the

total absorbance at the wave length of λ can be
expressed with the following equation:

A = ∑ An = b∑ εn cn
λ λ λ

n n

How do you measure the molar absortivity (ε) ?