Capillary Electrophoresis: Introduction, instrumentation, procedure, advantage and its application

Capillary Electrophoresis

 Introduction of Capillary Electrophoresis

The introduction of capillary zone electrophoresis is a major advance in serum protein electrophoresis. Referred as high-performance capillary electrophoresis (HPCE), capillary zone electrophoresis (CZE), Free solution capillary electrophoresis (FSCE), and Capillary electrophoresis (CE).

Instrumentation 

Instrument includes-Capillary outlet
Capillary inlet
Capillary tube
High voltage supply
Anode
cathode
Reservoir
Electrolyte buffer
Detector
Data acquisition as shown above image

Procedure of Capillary electrophoresis

  1. Fill the tube with the sample solution.
  2. Carry out the sample application by high voltage injection or by pressure injection.
  3. Apply a high voltage of 20kV.
  4. Two forces act on the proteins: The attraction of first the negatively charged proteins toward the positive electrode anode and second, an electro-osmotic force (EOF).
  5. EOF drags all the molecules towards the cathode irrespective of the charge.
  6. During this migration, positively charged molecules move faster than negatively charged ones. Detection of resolved molecules is done by a UV monitor and the signal is recording in a recorder.
  7. A capillary electrophoretogram can obtain by plotting the signal against time.

Advantages 

  • Automation
  • High speed
  • High voltage can be applied
  • Proper heat dissipation
  • A small amount of sample is enough

Applications of Capillary electrophoresis

  • Used to separate:
  • Inorganic ions
  • Amino acids
  • Organic acids
  • Drug
  • Vitamins
  • Porphyrins
  • Carbohydrates
  • Oligonucleotide
  • Proteins
  • DNA

Further Readings

  1. Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 5th Edition
  2. Principles and Techniques of Biochemistry and Molecular Biology, Wilson and Walker 7th Edition
  3. Short protocols in molecular biology, 4th edition
  4. Pulsed-field gel electrophoresis( A practical Guide )by Bruce Birren21.
  5. Molecular Biology. Editor: David Freifelder, 2nd ed, Publisher Jones and Bartlett Inc.
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