GUIDED BY :

PROF.RAHUL KADU SIR

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• Ultraviolet (UV) is electromagnetic radiation with
wavelength from 10 nm to 400 nm.
• It is shorter than that of visible light but longer than X-
rays.
• UV radiation is present in sunlight, and contributes
about 10% of the total electromagnetic radiation output
from the Sun.

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 Burning things to a crisp (as in the Bunsen
Spectroscope) can tell us about the atoms
contained in a molecule…
 But this doesn’t tell us anything about how the
molecule is put together
 Fortunately, certain molecules absorb light in a
characteristic way. Over the years, this has helped
us identify and quantify biological molecules.

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 Absorption occurs when the energy contained in a
photon is absorbed by an electron resulting in a
transition to an excited state.
 Absorption occurs when the energy contained in a
photon is absorbed by an electron resulting in a
transition to an excited state.

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 hν

Π π*

So, our absorption spectrum

should probably look like
this:But they don’t…

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This is because molecules are always rotating
and vibrating. Each rotational or vibrational
state slightly changes the energy of the
transition.
So the lineshape of our absorption spectra is
normally distributed

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•The absorption efficiency of an analyte is affected
by:

1.SOLVENT:The choice of solvent changes the peaks of

shorter and longer wavelength.
2.Concentration: Sample with higher concentration
changes the position of absorption bands and do not follow
Beer’s Lambert Law.
3.Temperature: Change in temperature causes
various impacts like expansion and contraction of
solvent,changes in reaction rate etc.

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 A=ebc= log (I0 / IT)
e = Extinction coefficient

c = Concentration

b = Path length

Pierre Johan
Bouguer Lamber
(1698- t (1728-
1758) 1777)

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 Beer-Lambert Law
•The absorbance of light is directly proportional to the
thickness of the media through which the light is being
transmitted and the concentration of absorbing species;
that is, A = εbc Total cross section of
particles concentration

Absorption =

y
Cross section of beam

x
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UV/Visible Spectroscopy:
Instrumentation
#Instruments for measuring the absorption of U.V. or
visible radiation are made up of the following components;
1. Sources (UV and visible).
2. Filter or Monochromator.
3. Sample containers or sample cells.
4. Detector.
5.Amplifier.
6.Recorder.

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UV/Visible Spectroscopy: Light Sources

Xenon, Mercury/Xenon

Flash Arc-Lamps – light

generated from Xe
plasma
Pure Xenon has very wide emission spectrum 200-1200
nm
Xenon/Mercury is blue shifted for more power in the UV
region, used more often for sterilization

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 UV/Visible Spectroscopy: Light Sources

Deuterium

D2 gas is discharged by contact with a high voltage

tungsten cathode
Continuous spectrum from ~150 nm - ~370 nm
Usually used in conjunction with a tungsten/halogen
source, which handles the visible spectrum
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The detector

Silicon diode:
Basically a solar cell – light ionizes n-doped
(phosphate) silicon, placing the electrons in the
conduction band (i.e. having a voltage).

Wide wavelength range, less sensitivity

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Photomultiplier

In photomultipliers, light hits a photocathode, releasing a

small number of electrons, which are then made to
collide with a series of dynodes, each more positive than
the last

Each collision produces more and more activated

electrons
Sensitive, but noisy. Pretty much needed for low
energy (IR) photons

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UV absorption spectroscopy is one of
the best methods for determination of
impurities in organic molecules.
Benzene appears as a common impurity
in cyclohexane and can be easily
detected by its absorption at 255nm.
It may be used as a detector for HPLC.

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UV absorption spectroscopy can be used for
quantitative analysis of compounds that
absorb UV radiation.
Molecular weight of compounds can br
measured spectrophotometrically by
preparing the suitable derivatives of these
compounds.
Kinetics of reaction can also be studied
using UV spectroscopy. The UV radiation is
passed through the reaction cell and the
absorbance changes can be observed.

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 ADVANTAGES OF UV
1) The process only requires a minimal amount of electricity.
2) It is also very environmental friendly.
3) This is because it does not emit any by-products such as sodium into
the environment. Therefore, there are no chemicals handled.
4) The UV system is very cost effective.
5) The ultraviolet radiation method is one of the most used methods of
water softening concerning the removal of bacteria.
6) The ultraviolet rays are so powerful such that when they are
directed to the target water they instantly destroy all the pathogens
in the water.
7) Also, the system does not consume a lot of energy because only the
ultraviolet radiations are the ones needed.

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1) The water must be very clear so that the
Ultra Violet radiation can pass throug
and destroy the pathogens.
2) The process can occur in a place which
has no electricity.
3) Ultraviolet radiation method is only able
to deal with microorganisms, and that
means that if the hard water contains
minerals such as magnesium and
calcium, then the rays will not be able to
eliminate them.

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THANK YOU

BY-
AKSHAY NANGRE
MALHAR MANGLE
PARTH KULKARNI
AMOGH DAHITULE

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