Capillary Electrophoresis

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Transcript Capillary Electrophoresis

Capillary
Electrophoresis
An investigation of the theory
and operation, and its
application to environmental
and DNA analyses.
Kristina Schmit
Douglas Jones
Jordan Simon Fromholz
John Jr. Yoshihara

Topics of Discussion

What is Capillary Electrophoresis?
Theoretical Background

Capillary Electrophoresis System

Applications and Contributions

Environmental Applications
 Applications to DNA Analysis

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Conclusion
What is Capillary
Electrophoresis?
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Analytical separation technique
The “movement or migration of
ions… under the influence of an
electric field.”
Regularly applied to inorganic,
organic, environmental,
pharmaceutical, and biological
analyses
Theoretical Background


Separating trace amounts of ionic
species can be a highly difficult
and tedious process.
The separation process depends
on the ions having different
migration speeds. The velocity of
ion migration [m/s], is defined by::
ν = μeE
Theoretical Background
(cont.)

The mobility indicates how fast the
given ion or solute moves through a
medium.
q
e 

r

Electrical forces move the ion
forward, and frictional forces oppose
the motion. [1]
The Development of CE
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Old methods involved using
paper or gels to transport the
charged species to the
cathode or anode.
Long analysis coupled with
tedious lab time made
process almost impractical.
The addition of a fused silica
capillary in the 1980’s
created a more efficient,
precise method of separation
and identification.
The CE System
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The capillary serves two
functions
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High voltage is passed
through the tube charging
the molecules.
The tube also allows for the
flow of ions from one node
to the other.
Ions flow by means of an
induced current on the
walls of the capillary,
creating an ‘electron
driven pump’.
Ion Flow and Analysis
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Applying a voltage
causes the flow of metal
ions on the wall of the
capillary creating an
electroendosmotic flow
(EOF).
The induced flow
transports the
separated ions past an
ultra violet absorbance
that calculates their
physical properties.
Capillary Electrophoresis
Applications & Contributions
Environmental
Applications

CE is an important tool for
detecting and separating a
wide variety of compounds.

CE is used in environmental
monitoring.
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Example: Herbicides and
pesticides runoff polluting
streams & groundwater.
Environmental
Applications
 CE is used in determining amounts of
Inorganic and organic species in
Environmental water and sediments.
 Example: Toxic substances
methyl mercury in drinking water.
 CE is used in separating ions.
e.g. lead contamination in ground soil.
Environmental Impact
Cost Efficiencies
 CE uses less organic solvent;
less product waste = less harm
to environment.
 Shorter analysis time; reduces
Cost = more efficient.
 Easier, cleaner preparation.
= more convenient.
Applications to DNA
Analysis

Mutation detection

Provide for future advances in diagnosis
of human genetic diseases.

CE is capable of analyzing both known
and unknown mutations.

Quicker separation technique while not
sacrificing effectiveness or range of
applicability.
Applications to DNA
Analysis (cont.)

Human Genome Project
Lions share of separation and
analysis done by CE.
 Provides for rapid DNA analysis
and high throughput.

 Capillary
Array Electrophoresis
What This Means
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Separation technique capable of separating trace
amounts of ionic species.

Is a cleaner, cheaper, faster and more
environmentally favorable method of detecting
and separating environmental contaminants.
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Contributions to biological sciences have
provided foundation for profound future
advances.
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Future developments will broaden range of
applicability