In this slide contains Electrophoresis Principle and its Types. Presented by: KHALID KUWAITY (Department of pharmaceutical analysis). RIPER, anantapur

1. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 1
A seminar as a part of curricular requirements
for 1st year M. Pharm 1st semester
Presented by
Khalid Osman
Reg. no:20L81S0710
Under the guidance of
Dr. P. Ramalingam, Ph.D
Director – R&D Division
Electrophoresis Principle
and its types

2. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 2
• Introduction to electrophoresis
• Principles
• Types of electrophoresis
Contents

3. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 3
• Electrophoresis is a technique commonly used in the lab to separate
charged molecules, like DNA, according to size.
• is a separation method that is based on the migration of charged
species in a supporting medium (a liquid or a hydrophilic gel)
under the influence of an electric field
Introduction to electrophoresis

4. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 4

5. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 5
• is a general term that describes the migration and separation of
charged particles (ions) under the influence of an electric field. An
electrophoretic system consists of two electrodes of opposite charge
(anode, cathode), connected by a conducting medium called an
electrolyte.
• The separation effect on the ionic particles results from differences
in their velocity (v), which is the product of the particle's mobility
(m) and the field strength (E):
V=Me
Principles

6. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 6
• Paper electrophoresis
• Capillary electrophoresis
• Zone electrophoresis
• Iso electric focusing
• Gel electrophoresis
Types of electrophoresis

7. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 7
• The technique of paper electrophoresis is simple and inexpensive and
requires only micro quantities of plasma for separation
• The support medium is a filter paper
• The electrophoresis apparatus in its simplest form consists of two
troughs to contain buffer solution through which electric current is
passed
• Frequently used in isolating protiens, amino acids
Paper electrophoresis:

8. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 8
• CE is used to analyze and separate unknown samples based
on their mobility under an applied electrical voltage.
• Mobility is dependent upon the molecular charge, the atomic
radius, and viscosity.
• Ions are the species that migrate to opposing ends of an
applied voltage, and its motion is dependent on the applied
electric field.
• the sample ions migrate through the capillary at different speeds,
causing them to divide
Capillary electrophoresis

9. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 9
• is the most widely used mode. It is fundamentally the simplest form
of CE, mainly because the capillary is only filled with buffer
• Separation occurs because solutes migrate in discrete zones and at
different velocities and are separated solely based on differences
• Both anionic and cationic solutes can be separated
Zone electrophoresis:

10. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 10
• is a technique for separating charged molecules, usually proteins or
peptides, on the basis of their isoelectric point (pI), i.e., the pH at
which the molecule has no charge.
• IEF works because in an electric field molecules in a pH gradient
will migrate towards their pI.
Isoelectric focusing electrophoresis:

11. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 11
• is a technique commonly used in laboratories to separate charged
molecules like DNA, RNA and proteins according to their size.
• Charged molecules move through a gel when an electric current is
passed across it.
Gel electrophoresis:

12. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 12
• Mostovenko, E., Hassan, C., Rattke, J., Deelder, A. M., van
Veelen, P. A., Palmblad, M., 2013, Comparison of peptide and
protein fractionation methods in proteomics, EuPA Open
Proteomics, 1: 30-37.
• R.J. Fritsch, I. Krause, in Encyclopedia of Food Sciences and
Nutrition (Second Edition), 2003
References:

13. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 13