Module 5 : Analysis of biomolecules

Lecture 34 : Thin Layer Chromatography

Figure 34.1: Principle of thin layer chromatography.

Operation of the technique-Several steps are required to perform a thin layer chromatography to analyze a complex samples. These preparatory and operational steps are as follows: 

Thin Layer Chromatography Chamber- Thin layer chromatographjy chamber (rectangular or cylindrical) is made up of transparent non-reactive material, mostly glass (Figure 34.2). It is covered from top with a thick glass sheet and the joints  are sealed with a high vacuum grease to avoid loss of solvent vapor. All three sides of the chamber is covered with a whatman filter paper to uniformly equilibrate the chamber. A solvent system is filled in the chamber and it is allowed to humidify the chamber with the solvent vapor. It is important for uniform running of solvent front during TLC.