Thin Layer Chromatography
Aim: To separate a mixture of amino acids by Thin Layer Chromatography (TLC) and identifying the unknown amino acids in comparison with known amino acids by measuring their Rf values.
Principle: Thin Layer Chromatography (TLC) is a sensitive, fast, simple and in-expensive analytical technique. It is a solid-liquid form of chromatographic technique in which one phase is solid (or stationary phase) and another one is liquid (or moving phase). It involves spotting of the sample to be analyzed near one end of a sheet of glass or plastic that is coated with a thin layer of an adsorbent. The adsorbents most commonly used in chromatography are silica gel (SiO2 x H2O) and alumina (Al2O3 x H2O). Both of them are polar, but alumina is more. Silica is acidic whereas alumina is available in neutral, basic, or acidic forms.
The sheet is placed on end in a covered jar containing a layer of solvent (generally 1-2 cm layer from the bottom). As the solvent rises by capillary action up through the adsorbent, differential partitioning occurs between the components of the mixture dissolved in the solvent and the stationary adsorbent phase. The more strongly a given component of a mixture is adsorbed onto the stationary phase, the less time it will spend in the mobile phase and the more slowly it will migrate up the plate.
Kit Description:Cat. #BB-C030: This kit is designed to perform 10 experiments.
BioBharati Thin Layer Chromatography Teaching Kit contains 5 different standard amino acids stock solutions and different mixture of test solutions, TLC plates, TLC solvent, Ninhydrin solution are provided in this kit. The reagents are more than enough to carry out 10 experiments. BioBharati Thin Layer Chromatography Teaching Kit enables separation of mixture of different amino acids and their identification by measuring their respective Rf value.
Students will carry on the experiment as stated in “Kit procedure” and identify the amino acids by determining the Rf values.