TECHNIQUES OF ELECTROPHORESIS

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.

16. 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.

17. 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.

18. REFERENCE www.google.com

19. Q UESTIONS ?

20. Thank You