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Concept
Thin-layer chromatography
Summary
Thin-layer chromatography (TLC) is a chromatography technique that separates components in non-volatile mixtures.
It is performed on a TLC plate made up of a non-reactive solid coated with a thin layer of adsorbent material. This is called the stationary phase. The sample is deposited on the plate, which is eluted with a solvent or solvent mixture known as the mobile phase (or eluent). This solvent then moves up the plate via capillary action. As with all chromatography, some compounds are more attracted to the mobile phase, while others are more attracted to the stationary phase. Therefore, different compounds move up the TLC plate at different speeds and become separated. To visualize colourless compounds, the plate is viewed under UV light or is stained. Testing different stationary and mobile phases is often necessary to obtain well-defined and separated spots.
TLC is quick, simple, and gives high sensitivity for a relatively low cost. It can monitor reaction progress, identify compounds in a mixture, determine purity, or purify small amounts of compound.
The process for TLC is similar to paper chromatography but provides faster runs, better separations, and the choice between different stationary phases. Plates can be labelled before or after the chromatography process with a pencil or other implement that will not interfere with the process.
There are four main stages to running a thin-layer chromatography plate:
Plate preparation: Using a capillary tube, a small amount of a concentrated solution of the sample is deposited near the bottom edge of a TLC plate. The solvent is allowed to completely evaporate before the next step. A vacuum chamber may be necessary for non-volatile solvents. To make sure there is sufficient compound to obtain a visible result, the spotting procedure can be repeated. Depending on the application, multiple different samples may be placed in a row the same distance from the bottom edge; each sample will move up the plate in its own "lane.
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