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Introduction to Soil Chemistry

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alization of components on a thin-layer plate. Detection methods similar <strong>to</strong><br />

those used in HPLC are used in capillary electrophoresis. The type of separation,<br />

ion, organic ion, or ionic biomolecule will determine which detection<br />

method is best.<br />

9.5.5. Electrophoresis Applied <strong>to</strong> <strong>Soil</strong><br />

Gel electrophoresis has been applied <strong>to</strong> soil DNA and RNA extracts using<br />

procedures similar <strong>to</strong> those used in DNA testing used for forensic analysis.<br />

Capillary electrophoresis has also been applied <strong>to</strong> the analysis of ionic species<br />

extracted form soil. While these processes show promise for the elucidation<br />

of valuable information about soil, neither is used for common, routine soil<br />

analysis [11–13].<br />

9.6. IDENTIFICATION OF COMPOUNDS SEPARATED BY<br />

CHROMATOGRAPHIC PROCEDURES<br />

There are three ways of “identifying” compounds once they are separated.The<br />

simplest is that for which chroma<strong>to</strong>graphy is named. If a mixture of colored<br />

compounds is separated, and then where and when they elute or are found on<br />

a thin layer or gel is identified by searching for the color. After separating and<br />

visualizing a colorless component, it must be identified. This can be done in<br />

one of two ways. The first and simplest is by R f or R t, which are the distance,<br />

relative <strong>to</strong> some fixed point, that compounds move during a chroma<strong>to</strong>graphic<br />

procedure or between the time they enter and exit the chroma<strong>to</strong>graphic<br />

column [see equation (9.1) and Table 9.4 for further details on the use of R f<br />

and R t]. The formulas for R f and R t are:<br />

R<br />

R<br />

f<br />

t<br />

identification of compounds 189<br />

distance spot moved<br />

=<br />

distance elutant moved<br />

= Time from injection <strong>to</strong> <strong>to</strong>p of peak<br />

Table 9.4. The R f Values of Spots on Thin-Layer Sheets<br />

Shown in Figure 9.6 a<br />

Spot Calculation Rf A1 3/15 0.20<br />

A2 9/15 0.60<br />

B1 2/15 0.13<br />

B2 9/15 0.60<br />

B3 12/15 0.80<br />

a The distance that the solvent front moved is 15cm; therefore, this<br />

is the denomina<strong>to</strong>r for all calculations.<br />

(9.1)

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