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MOVING BOUNDARY ELECTROPHORESIS . pptx

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MOVING BOUNDARY ELECTROPHORESIS . pptx

  1. 1. Presented by Roshan Yadav M.pharm(Pharmaceutics). Submitted to Dr. Yele Vidyasrilekha. MOVING BOUNDARY ELECTROPHORESIS
  2. 2. Contents • Introduction • Principle of Electrophoresis • Types of Electrophoresis • Moving Boundary electrophoresis • Instrumentation • Advantages and Disadvantages • Application
  3. 3. Introduction • Electrophoresis is defined as the migration of charged particle through a solution under the influence of an external electric feild. • Ions that are suspended between two electrodes tends to travel towards the electrodes that posses opposite charge.
  4. 4. Principle • The fundamental principle of electrophoresis is that any charged ions or molecules migrate towards oppositely charged electrode under the influence of external electric field. • The rate of migration of these ions depends on its charge size and the applied electric field.
  5. 5. Types of electrophoresis 1)zone electrophoresis a) Paper electrophoresis b) Gel electrophoresis c) Thin layer electrophoresis 2) Moving boundary electrophoresis a) Capillary b) Isotachoporesis c) Isoelectric electrophoresis d) Immuno electrophoresis
  6. 6. Moving boundary electrophoresis • Moving boundary electrophoresis is technique for separation of chemical compounds by electrophoresis in a free solution. • Moving boundary electrophoresis is developed by Tiselius. • The principle of moving boundary electrophoresis is that there is motion of charge particle through stationary liquid under the influence of an electric field.
  7. 7. Instrumentation • Apparatus consist of “U” tube with electrode located at the two ends used to apply on electric field. • The lower part of the cells is filled with lyophilic solution under examination sometimes the sample solution is introduced into the bottom of the “U” tube through capillary arm while upper part contains only the buffer solution. • Care must be taken to minimize the disturbing effect of convection caused by an increase in temperature during the passage of current through the solution. • The apparatus is placed in constant temperature .
  8. 8. Advantages • Biologically active fraction can be removed without the use of denaturing agent. • Minute concentration of sample can be detected. • A reference method for measuring electrophoretic mobility.
  9. 9. Disadvantages • Expensive. • Resolution of the techniques is low due to overlapping of the sample component.
  10. 10. Applications • To separate proteins and peptides. • For research in enzymology, immunology. • Used to quantitative analysis of complex mixture of macromolecules.
  11. 11. THANK YOU

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