CONTENT Capillary Electrophoresis 1) INSTRUMENTATION 2) APPLICATION High voltage electrophoresis 1) INSTRUMENTATION 2) APPLICATION Low voltage electrophoresis 1) INSTRUMENTATION 2) APPLICATION

1. TECHNIQUES OF ELECTROPHORESIS
Presented by :Yogesh Mhadgut
M.Sc. Part 02
Semester : 04

2. CONTENT
 Capillary Electrophoresis
1) INSTRUMENTATION
2) APPLICATION
 High voltage electrophoresis
1) INSTRUMENTATION
2) APPLICATION
 Low voltage electrophoresis
1) INSTRUMENTATION
2) APPLICATION

3. CAPILLARY ELECTROPHORESIS

4. PRINCIPLE OF CE
• It is an analytical technique.
• CE separates ions based on their electrophoretic
mobility with the help of an applied voltage.
• The electrophoretic mobility depends upon the
charge on the molecule, viscosity and the atom’s
radius.
• The rate of particle moving is directly
proportional to the applied electric field

5. Neutral species is not affected, only ions moves with
the electric field.
If 2 ions are the same size, the one with greater
charge move fastest.
CE provides high resolution separation.
It is useful technique because there is a large range
of detection methods available.

6. INSTRUMENTATION OF CE
• A typical CE system consists of a :
• Buffer solution (like NaH2PO4).
• High voltage power supply
• Sample injection
• A capillary tube with internal diameter of 10-100mm
and 20-100cm length.
• Detector
• Output device

7. Sample injection
• Hydrodynamic injection: By applying
pressure, by applying vacuum.
• Electro kinetic injection: By using electric
supply.
DETECTOR
• Similar to GC,HPLC.

8. APPLICATION OF CE
• Genetic analysis
• Analysis of pharmaceuticals
• Protein characterization

9. LOW VOLTAGE ELECTROPHORESIS
The apparatus commonly used for LVE is of 2
types:
• HORIZONTAL STRIP
• VERTICAL STRIP

10. HORIZONTAL STRIP

11. VERTICAL STRIP

12. • Here the essential 2 compartments to hold the
buffer and electrodes and a suitable carrier for
support medium.
• The end of support medium are in contact with
buffer compartment.
• A design of carrier depends on the medium.
• To provide voltage gradient 5 Vcm-1.

13. APPLICATIONS OF LVE
• To separate any ionic substance.
• Separation of sugars.
• The examination of biological and clinical
specimens for amino acids and proteins.

14. HIGH VOLTAGE ELECTROPHORESIS
The apparatus commonly used for HVE is of 2
types:
• ENCLOSED STRIP
• IMMERSED STRIP

15. ENCLOSED STRIP
• In principle, paper electrophoresis is simpler than
paper chromatography.
• There are 2 features of an ion which are most
important to its electrophoretic behaviour; the size of
the ion and the size of the charge it carries.
• These feature opposite to each other; the bigger the
size the slower the ion will travel; the bigger the charge
the faster it will go.

17. IMMERSED STRIP METHOD
WORKING:
• In this method the sample is applied to a strip of
filter paper maintained with buffer solution and
the strip is immersed in a large volume of
buffer-immiscible, electrically insulating and
volatile cooling liquid.
• Apparatus can also be designed to accumulate
cooling liquids which have a higher density than
that of buffer solution.

18. • To obtain voltage gradients up to 100 Vcm-1, high
voltage and current supplying.
• Using cooling plates for heat dissipation
generated by high voltage.
• Less than of 1 h time required.

19. REFERENCE
• www.google.com

20. QUESTIONS?

21. Thank You