ISHA RAJPUT (19)

MSc II SEM
INSTRUMENTATION ASSIGNMENT

• UV- VISIBLE SPECTROSCOPY

•
INTRODUCTION
# Who invented- Arnold Beckman, 1941
# UV-Visible spectroscopy is a type of absorption spectroscopy, deal with
the use of the electromagnetic spectrum for the analysis of different
compounds by observing and understanding the interaction of this
spectrum with the compounds.
# This technique is based on the absorption of radiation in the UV-Visible
region and the emission of energy from the electrons when they travel
from the excited state to ground state.
# Purpose-Quantitative and qualitative determination of analyte.
#Ranges:-
UV-200-400nm
VISIBLE-400-800nm
# U-V visible absorption spectra originate from electronic transition
within a molecule
■ These transitions involving promotion of valence electrons from the
ground state to the higher-energy state(excited state) are called electronic
excitations and are caused by the absorption of radiation energy in the
UV-visible regions of the electromagnetic spectrum.
PRINCIPLE OF UV-VISIBLE:-
The Principle of UV-Visible Spectroscopy is based on the absorption of ultraviolet light or
visible light by chemical compounds, which results in the production of distinct spectra.
ISHA RAJPUT (19)
MSc II SEM
INSTRUMENTATION ASSIGNMENT
Beer- Lambert law:- As per the Beer-Lambert law, the greater the number of absorbing
molecules (that have the ability to absorb light of a specific wavelength), the greater the extent
of absorption of the radiation.

ELECTRONIC TRANSITION
According to molecular orbital theory, the excitation of a molecule by the absorption of
radiation in the UV-visible regions involves promotion of its electrons from a bonding, or non
bonding (n)orbital to an antibonding orbital.
■o.o* transition ■n-o* transition
■ π-π* transition ■n-π* transition
ISHA RAJPUT (19)
MSc II SEM
INSTRUMENTATION ASSIGNMENT
ABSORPTION AND INTENSITY SHIFT

INSTRUMENTATION

COMPONENTS:
1. Radiation Source
# A stable continuous radiation is required.
ISHA RAJPUT (19)
MSc II SEM
INSTRUMENTATION ASSIGNMENT
# It must be of the sufficient intensity for the transmitted energy to be detected at the end of the optical path.

Deuterium Lamp (deuterium gas).

-UV Region
-Wavelength Range: 190-420nm
Tungsten Lamp (halogen- iodine and bromide)
–Wavelength Range: Part of the UV and Visible
Xenon Lamp (xenon gas)
–Wavelength Range: 190-800nm

2. Monochromator
# It is a device that can we prymatic radiation into component wavelengths.
# The monochromator unit consists of:

Entrance slit – defines narrow beam of radiation from source.

Collimating lens (polished surface) – collimates the lights.
ISHA RAJPUT (19)
MSc II SEM
INSTRUMENTATION ASSIGNMENT
Prism (make-quartz)- disperses the light into specific wavelength.
Focusing lens –captures the dispersed light & sharpens the same to the sample cuvette via exit
slit
Exit slit- allows the corrected wavelength of light to the sample cuvette.
Filters – it limit the radiation to be absorbed by a sample.
3. Sample Holder
 The cells or cuvettes are used for handling liquid samples.
 For study in UV region, the cells are prepared from quartz or fused silica whereas colour
corrected fused glass is used for visible region.
 Glass is suitable for visible but not for UV spectroscopy because it absorbs UV radiation.
 Cleaning is carried out washing with distilled water or with dilute alcohol, acetone.

4. Detector :-
 Device which converts light energy into electrical signals, that are displayed on readout
devices.
 The transmitted radiation falls on the detector which determines the intensity of radiation
absorbed by sample.
 Requirements of an ideal detector:
✓It should give quantitative response.
✓ should have high sensitivity and low noise level.
✓It should have a short response time.
TYPES OF DETECTORS
ISHA RAJPUT (19)
MSc II SEM
INSTRUMENTATION ASSIGNMENT
1. Barrier layer cell/Photovoltaic cell
2. Phototubes/Photo emissive cell
3. Photomultiplier tube

5. Amplifier and Recorder :-

 Amplifier amplifies signal coming from detector
 Recorder records them which is displayed on readout device.

TYPES OF UV- VISIBLE SPECTROMETERS

1. Single Beam spectrometers

As the name suggests, these instruments contain a single beam of light. The
same beam is used for reading the absorption of the sample as well as the
reference. The radiation from the source is passed through a filter or a suitable
monochromator to get a band or a monochromatic radiation. It is then passed through the
sample (or the reference) and the transmitted radiation is detected by the photodetector.
The signal so obtained is sent as a read out or is recorded.

2. Double Beam spectrometer

In a double Beam spectrometer, the radiation coming from the monochromator is split
into two beams with the help of a beam splitter. These are passed simultaneously
through the reference and the sample cell. The transmitted radiation are detected by
the detectors and the difference in the signal at all the wavelength is suitably amplified
and send to the output.
ISHA RAJPUT (19)
MSc II SEM
INSTRUMENTATION ASSIGNMENT

ADVANTAGES

 Quick and accurate.

 Less interference
 Less volume of sample.
 Various class of compound detection.

DISADVANTAGES

 Chromophore dependent
 Liquid samples only.
 Stray light headache
 Specificity at low wavelength