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Author Topic: THIN LAYER CHROMATOGRAPHIC TECHNIQUES- Preparation of TLC plates  (Read 16268 times)
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Francis Umeoguaju
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« on: April 10, 2010, 07:51:13 am »


THIN LAYER CHROMATOGRAPHIC TECHNIQUE

Thin layer chromatographic (TLC) is a type of chromatography which involves the coating of a adsorbent on glass, plastic plates or metal foil. This is used for separating components such as sugars, amino acids, liquids, drugs etc.

The separation of sample into their various components by TCL is based on the differences in the distribution coefficient kd, of the component of a sample in the mobile phase. As a result of the differential solubility, the rate of migration of the components of a sample in a mobile phase will differ; substances that are very soluble in the solvent tend to move close to the solvent front while those less soluble will have lesser tendency to be lifted up.

Preparation of TLC plates: There are different types of adsorbent material or stationary phase used to prepare TLC plates some commonly used adsorbent include silica gel, kelsigur and alumina. The choice of adsorbent depends largely on the class of compounds to be separated but silica gel is usually used for separating a wide variety of substance.

Preparation of slurry
; Silica gel is  weighed and is mixed with calcium sulphate Binder in the ration 17:3. The slurry is prepared by adding 1.5 to 2 parts of distilled water or buffer to 1 part of silica adsorbent and the resultant mixture is stirred properly.

Coating the plate; There are various technique used in coating with the slurry. These include; pouring immersion, spreading and spraying. The method I often use is that of spreading.

3 glass plates of dimension 15cm x 20cm are lined in a row on a white foolscap sheet spacer of about 1mm in used to line the sides. The slurry is then poured at the origin the centre plate and a glass rod is used to roll the slurry smoothly over the glass plate. After about 10mins, the spacers are then removed and the plates are air-dried for about 45 minutes.

The glass plates were then heated in an oven set at 110c for about 30mins, this treatment help activates the adsorbent.
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