Thin-Layer Chromatography(TLC)

TLC is used to separate and isolate low volatile compounds up to 1g.
2-5 components can be separated due to limited zone capacity. The sample capacity can increase by increasing the length and thickness of the layer.

 

The broad particle size range(25 um) contributes to the low efficiency of TLC, to increase the efficiency, the tapered layers is made.

- a negative velocity gradient in the direction of mobile phase is formed 

- the lower portion of a zone moves faster than the upper portion, so each component is focused as a narrow band.

Next we will have a look at sample application

- the sample 5-10% (w/v) solution in a volatile organic solvent. For example, a silica layer 1.0 mm thick is applied to 5 mg/cm2. 

- A margin of 1-2 cm is frequently left at both vertical edges of the layer to minimize uneven development.

- the sample is introduced to the layer by using manual syringe or pipetter.

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