More info about capillary electrophoresis
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Usually, the capillary is filled with an appropriate buffer. The
sample solution is injected from the inlet end of the capillary,
electric field is applied to the capillary ends and the components
migrate to the detector point with different velocities due to the
differences in their electrophoretic properties. Consequently, they
reach detector at different times. Results are present as a sequence
of peaks, where each peak represents ideally one component. Peak
area or peak height indicates the initial concentration of the component
in the mixture.
The main parameter, which provides electrophoretic differentiation
of the components in CE, is their charge-to-mass (e/m) ratio. Since
this ratio is highly characteristic, CE separation shows a very
high resolution and is suitable for the analysis of almost all classes
of compounds. The presence of ionisable groups ( - Si - OH) on the
inner surface of the quartz capillary gives rise to a phenomenon
called electroosmotic flow, which is just a bulk flow of the whole
liquid inside the capillary, occurring upon applying the voltage.
Electroosmotic flow enables simultaneous determination of positive
and negative charged compounds in one run. Even neutral compounds
can be analysed in one run together with charged species provided
a surfactant (e.g. sodium dodecyl sulphate) is added to the buffer
to form a micellar phase. This mode of CE, called micellar electrokinetic
chromatography is a very successful combination of electrophoretic
and chromatographic methods in which separation of compounds is
based on the differences in their charge-to-mass ratio and hydrophobic/hydrophilic
properties. Several other modes of CE can be realised on Capel instruments
regarding the nature of separated solutes and general goals of the
analysis. They are e.g. capillary gel electrophoresis, capillary
isoelectric focusing and capillary electrochromatography.
In the most of cases UV lamp is used as a source of the light
at the detector point. Monochromator, placed on the most upgraded
Capel-105M CE system, gives the highest flexibility to the user
enabling selection of the most appropriate wavelength for the detection
and spectral scanning of the separated compounds. Compounds which
do not absorb light in the UV range (e.g. inorganic ions), can be
analysed in the indirect mode with a certain UV - absorbing additive
in the buffer.
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