Friday, April 2, 2021

Difference between TLC and Paper Chromatography

The major difference between TLC and paper chromatography is that the TLC uses the stationary phase as alumina, silica, and its compositions, whereas in paper chromatography, specialized cellulose paper is used as a stationary phase to separate the analytes.
 
TLC and paper chromatography both are separation techniques of chromatography, which are used to separate the components or biomolecules such as carbohydrates, amino acids, and proteins. As a stationary phase, the paper chromatography uses cellulose paper, and the separation mechanism depends on the solid-liquid adsorption. The same adsorption mechanism uses in thin layer chromatography, depending on solubility in the mobile phase the molecules or components are separated on the stationary phase.

Particle separation in both paper and thin-layer chromatography is based on the polarity of the molecules on the stationary and mobile phase. After the chromatograph is completed, the different spots will appear, and the Rf factor will be determined.

Thin Layer Chromatography:

TLC is a separation technique of chromatography, which works based on the solid-liquid adsorption of compounds. It has a stationary phase of silica gel or alumina and a solvent in the form of a mobile phase, where the particles are separated based on their polarity towards both phases. In TLC the colored compounds appear as colored spots and colorless compounds are seen under UV light (UV cabinet) or using a spraying solution (Ninhydrin).

Paper Chromatography:

Paper chromatography is a separation technique used to separate compounds based on their liquid-liquid adsorption and solubility of the compound. The paper acts as a stationary phase and the mobile phase is liquid, and the particles are isolated according to their polarity toward both phases. In paper chromatography, the colored compounds appear as colored spots and colorless analytes are seen under using a spraying solution (Ninhydrin).

The key differences between thin layer chromatography (TLC) and paper are as follows:

  • TLC is based on the principle of "adsorption," whereas paper chromatography is based on the principle of "partition".
  • TLC is based on solid-liquid absorption whereas paper chromatography is based on solid-liquid absorption.
  • The glass plates coated with silica gel acts as the stationary phase in TLC whereas the water trapped in the cellulose filter paper acts as a stationary phase of paper chromatography.
  • Thin-layer chromatography takes more time in preparation, whereas paper chromatography takes less preparation.
  • TLC allows the use of corrosive reagents, while paper chromatography does not because the corrosive agents may damage the paper. This is due to the TLC plate is resistant to strong solvents and color-creating agents.
  • Paper chromatography rapidly separates the molecules whereas thin layer chromatography requires more time.
  • In TLC the individual spots of solutes can be seen under UV-lamp whereas in paper chromatography cannot be evaluated under the UV light.
  • The development of chromatogram takes place (flow of mobile phase) in paper chromatography can ascending, descending, radial, and two-dimensional whereas the flow of mobile phase in TLC chromatogram develop in ascending order.
 
  
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