Analytical Techniques

P R E S E N T E D T O : M A’ A M S A D I A
P R E S E N T E D B Y: K A I N AT M U N I B A
ASSIGNMENT 1
9 T H . S E P. 2 0 2 0
Analytical techniques
Commonly used
Liquid chromatography
Gas chromatography
Mass spectroscopy
UV/IR/VIS spectroscopy
Spectrophotometry
Atomic absorption
(R. Sandford, P.J. Worsfold, 2005)
Visible and ultraviolet spectroscopy
Ultraviolet (UV)-visible spectroscopy is a type of
absorption spectroscopy in which UV-visible light is absorbed by the
molecule. Absorption of the UV-visible radiations results in the
excitation of the electrons from lower to higher energy levels.
Working:

UV visible spectroscopy is related to the interaction of light with matter.

As light is absorbed by matter, the result is an increase in the energy content of the atoms or
molecules.
When ultraviolet radiations are absorbed, this results in the excitation of the electrons from the
ground state towards a higher energy state.
Molecules containing π-electrons or non-bonding electrons (n-electrons) can absorb energy in the
form of ultraviolet light to excite these electrons to higher anti-bonding molecular orbitals.
The absorption of ultraviolet light by a chemical compound will produce a distinct spectrum which
aids in the identification of the compound.

(R. Jyotika, J.Gagandeep, 2019)

Instrumentation of UV Spectroscopy
Light source
Monochromater
Sample solution reference cells
Detector
Amplifier
Recording devices
Advantages
It requires no physical contact between sample and the instruement.
The instrument can be positioned directly on the sample(like skin) or kilometres
away(like satellites)
No sample preparation
Quantitative results
Accuracy of results
Simple to use
Disadvantages
Time taken to prepare the instrument
Clear the area of any outside light, electronic noise, or other outside
contaminants can interfere with the spectrometer's reading.
Even a small bit of outside light or vibration from a small electronic device could
interfere with the results
C. Maria., 2018
 Application
1. Detection of Impurities
2. Structure elucidation of organic compounds
3. To detect the presence or absence of functional group in the compound
4. Kinetics of reaction can also be studied
5. Many drugs are either in the form of raw material or in the form of
formulation. They can be assayed by making a suitable solution of the drug in a
solvent and measuring the absorbance at specific wavelength. 
6. Molecular weights of compounds can be measured spectrophotometrically by
preparing the suitable derivatives of these compounds.
7. UV spectrophotometer may be used as a detector for HPLC. (M. Tomislav., 2018)
Atomic absorption spectrophotometer
It was introduced or analytical purpose by Walsh, Alkemade and Milatz in 1956.
It is an absorption spectroscopic method in which radiation from a source is
absorbed by non-excited atoms in the vapour state.
It deals with the absorption of specific wavelength of radiation by neutral atoms
in the ground state.
Principle
Basic Principle

Light from special monochromatic sources with the wavelength of the resonance line of the
element to be determined is passed through a vapor of neutral atoms.
The diminution of the light beam by the activated atoms is monitored by a detector and
recorded by a read-out system.
Because the wavelength of the light emitted from the excited atoms in the flame is identical
with the wavelength of the source line, radiation from both, the lamp and the flame, is passed
by the monochromator into the light detector
Instrumentation
Sharp-line radiation source (a hollow cathode lamp)
A solution nebulizer and burner
Monochromator
Photomultiplier
Recording system
 Advantages
High selectivity and sensitivity
Fast and simple working
Doesn’t need metal separation
Provides a reducing environment to be easily oxidized
Smaller quantities of sample (5-50 micro-Litre)
(D. L. Fry, 1956)
Disadvantages

Expensive
High operator skill
Individual source lamp is required for each element
Analysis is not simultaneous
Sample should be in solution form or at least volatile
Limited types of cathode lamp as they are expensive
 Applications
Qualitative and quantitative analysis
Determination of metallic elements in biological systems
Determination of metallic elements in food industry
Determination of Ca, Mg, Na, K in serum
Determination of lead in petrol
Determination of metals at trace level in solutions
Most common
Most common is Atomic absorption.
Thank You