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Electrophoresis
1. M.Sc.MLT
SEPERATION TECHNIQUES AND PHARMACEUTICAL CHEMISTRY
SUB CODE: 17PML4MC02
Prof incharge: Dr.Robert Selvam
presented by
A VAMSI KUMAR
Dep No: 17PML-014
Loyola College
2. INTRODUCTION
1. Theory of Electrophorosis.
2. principle of electrophorosis.
3. Types of electrophorosis.
a) Moving boundary electrophorosis
b) Zone electrophosis
3. Meaning of Electrophoresis:
• The term electrophoresis
describes the migration of a
charged particle under the
influence of electric field
( particle and
).
• Many important biological
molecules such as amino acids,
peptides, proteins, nucleotides,
nucleic acids possess ionisable
groups and, therefore, at any
given pH, exists in solution as
electrically charged species
either as cations or anions.
electrocharged
phoresismovement
4. Mol seiving:a crystalline substance (especially a zeolite) with pores
of molecular dimensions which permit the passage of molecules
below a certain size.
5.
6.
7.
8. Mol seiving:a crystalline substance (especially a zeolite) with pores
of molecular dimensions which permit the passage of molecules
below a certain size.
9. Classification of Electrophoresis:
• All modern electrophoretic apparatus have supporting media.
• A supporting medium is a physical support through which the
charged molecules get separated.
• It has two primary functions:
1. Adsorption and
2. Molecular sieving of the taken molecules which
are intended to be separated.
Now a question arises why to take a supporting
medium and why not to follow the basic principles
of electrochemistry, using an electrophoretic tank
with two electrodes for separating these charged
molecules. This is what had been done in the
beginning days.
10. Arne Wilhelm Kaurin Tiselius (10 August 1902 – 29
October 1971) was a Swedish biochemist who won
the Nobel Prize in Chemistry in 1948 "for his
research on electrophoresis and adsorption
analysis, especially for his discoveries concerning
the complex nature of the serum proteins."
11.
12. • The pioneering work on
electrophoresis by
was
performed in
• However, it was soon realized
that many of the problems
associated with this approach,
particularly the adverse effects
of convection currents (Current
caused by the expansion of a
liquid, solid, or gas as its
temperature rises.
Tiselieus et
al. in the year 1948
free solution only.
13. • This coventionl currents can be minimized by stabilizing the medium.
• This was achieved by carrying out electrophoresis on a porous mechanical support.
• Due to its viscosity (which is absent in free solution) the supporting media cut down
the convention currents due to which we get the separated molecules in sharp zones.
• Some of the frequently used supporting media are
1) Starch,
2) Agar,
3) polyacrylamide and
4) agarose.
ØDepending upon the presence or absence of supporting media the electrophoresis
can be classified as
I. Free electrophoresis and
II. Zone electrophoresis.
