Introduction to Soil Chemistry
Introduction to Soil Chemistry
Introduction to Soil Chemistry
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188 chroma<strong>to</strong>graphy<br />
9.5. ELECTROPHORESIS<br />
As with other chroma<strong>to</strong>graphic methods, there are an number of electrophoretic<br />
methods, including paper, gel, and capillary. Electrophoresis uses<br />
an electric current <strong>to</strong> move ionic species, either simple ions, amino acids, or<br />
complex proteins, through a medium or a capillary [i.e., capillary electrophoresis<br />
(CE)]. During this process, the ionic species typically move at different rates<br />
and are thus separated (CE is somewhat different, as discussed below).<br />
9.5.1. Sample Application<br />
In paper or gel electrophoresis, a sample may be applied with a syringe or a<br />
micropipette similar <strong>to</strong> the application of samples <strong>to</strong> thin layer plates. In some<br />
cases there may be “wells” in the gel that accept the solution containing the<br />
species <strong>to</strong> be separated. In capillary electrophoresis samples may be applied<br />
using electromigration, hydrostatic, or pneumatic injection.<br />
In all cases the ions <strong>to</strong> be separated must be soluble in and compatible with<br />
the stationary phases and buffers used.<br />
9.5.2. Movement of Species<br />
For electrophoresis the paper or gel is saturated with the required buffer at<br />
the desired pH. The ends of the paper or gel are placed in a buffer reservoir<br />
that contains the buffer with which the paper or gel is saturated and that also<br />
have electrodes connecting one end <strong>to</strong> the positive DC terminal and the other<br />
<strong>to</strong> the negative terminal of the power source. It is the electrical current that<br />
causes the movement of ionic species through the medium.<br />
In capillary electrophoresis a high voltage is used <strong>to</strong> produce electroosmotic<br />
flow, and both electricity and the buffer flow through the capillary, with the<br />
buffer flowing <strong>to</strong>ward the cathode. Both carry the sample through the capillary,<br />
and because of the flow of the buffer, both charged and uncharged species<br />
are separated.<br />
9.5.3. Stationary Phases<br />
Electrophoresis can be carried out using paper or a gel as the supporting<br />
medium. Typically it can be carried out only in media compatible with water<br />
since buffers or salt solutions are required <strong>to</strong> carry the electric current<br />
required for separation. Capillary electrophoresis is carried out in a fusedsilica<br />
capillary filled with buffer.<br />
9.5.4. Detection<br />
Once an electrophoretic separation has been accomplished, the paper or gel<br />
is sprayed or dipped in a visualizing solution similar <strong>to</strong> that used in the visu-