Capillary Electrophoresis

By: Waepril Chua

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 Outline
• Introduction/Background of Capillary Electrophoresis (CE)

• Instrumentation Setup

• Electroosmotic Flow (EOF)

• Data Collection

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 Capillary Electrophoresis
• Capillary electrophoresis
- technique used to separate ions or solutes using capillary tubes, which are about 25 to 100
µm in internal diameter.
- separation occurs due to differences in the migration velocity of different solutes.
v=µeE where E=electric field, µe=electrophoretic mobility
- velocity is proportional to the electric field
- electrophoretic mobility- measure of how fast an analyte will travel in a given
buffer/medium
µe = ___q__ where q=charge of the ion, r=ionic radius,
6 πηr η= solution viscosity

Skoog, D.A.; Holler, F.J.; Crouch, S.R. Principles of Instrumental Analysis, 6th ed; Thomson Brooks/Cole Publishing: Belmont, CA, 2007;
Chapter 30.
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 Instrumentation
• Two buffer reservoirs containing an anode
and a cathode

• The ends of capillaries are placed in the

two buffer reservoirs.

• To inject sample, replace one buffer

reservoir (normally the anode end) with a
buffer reservoir containing the sample.

• Voltage is applied to separate the

analytes.

Frazier, R.A.; Ames, J.M.; Nursten, H.E. Capillary electrophoresis for food analysis: method development; The Royal Society of Chemistry:
Cambridge, UK., 2000; Chapter 1.
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 Electroosmetic flow (EOF)
• The flow of liquid solution through the
capillary
• Double layer charge distribution (Stern’s
model)
• Rigid layer
- a layer of positive charges
- forms due to the excess of negative
charges in the inner surface of the
capillary
• Diffuse layer
- a layer of a mixture of positive and
negative ions
- diffusion of ions occurs due to thermal
motion

Frazier, R.A.; Ames, J.M.; Nursten, H.E. Capillary electrophoresis for food analysis: method development; The Royal Society of Chemistry:
Cambridge, UK., 2000; Chapter 1.
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 Collecting data
• An electropherogram is obtained. (Similar
to a chromatogram)

• Measure of detector response vs

migration time

• Flat profile

• Broadening is minimized.

• High separation efficiencies

Frazier, R.A.; Ames, J.M.; Nursten, H.E. Capillary electrophoresis for food analysis: method development; The Royal Society of Chemistry:
Cambridge, UK., 2000; Chapter 1.
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