Thin layer chromatography (TLC) is a chromatographic technique used to separate the Here is a popular method of producing TLC pipettes.‎Retention Factor · ‎Apparatus · ‎Spotting and Developing · ‎Common Problems in TLC. Thin layer chromatography, or TLC, is a method for analyzing mixtures by separating TLC is a sensitive technique - microgram ( g) quantities can be. Thin-layer chromatography (TLC) is a chromatography technique used to separate non-volatile mixtures. Thin-layer chromatography is performed on a sheet of glass, plastic, or aluminium foil, which is coated with a thin layer of adsorbent material, usually silica gel, aluminium oxide (alumina), or cellulose.‎Plate preparation · ‎Technique · ‎Analysis · ‎Applications.


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In this example, the triacylglyceride content of bat sebaceous glands was studied.

The lipid surface fraction was first separated by polarity on a TLC plate. The triacylglyceride band was then removed from the thin layer chromatography method with a spatula. The silica powder was transferred to a microcentrifuge tube with solvent.

After centrifugation, the stationary phase was left at the bottom of the tube, while the compounds remained dissolved in the solvent.

The triacylglycerides were then further separated by another physical property. In this case, the thin layer chromatography method dimension of separation was molecular size. TLC can also be used to monitor the progression of a chemical reaction.

Thin-layer chromatography - Wikipedia

In this example, the starting material thin layer chromatography method the reaction was used as a standard, and ran alongside the reaction solution on a TLC plate. This process was repeated at specific intervals over the course of the reaction.

As the reaction progressed, the starting material band diminished and the product band enlarged. When there was no change in bands, or all of the starting material was consumed, the reaction was complete.

Thin-layer chromatography

Finally, TLC plates can be used in bioassays. In this example, compounds were separated from red clover with TLC. Thin layer chromatography method band was then placed on bacteria growing on agar plates. Molecules that showed inhibited bacterial growth were further analyzed for their antimicrobial properties.

You should now understand the underlying theory of the separation, how to choose an appropriate mobile phase for your experiment, and how to set up and operate a TLC plate. An unknown compound 'A' may be compared to known standards 'B' through 'E'.

Schematic of a TLC plate. The retardation factor Rf should be consistent between thin layer chromatography method as long as the conditions are held constant.

Performing 1D Thin Layer Chromatography

Eluotropic Series for Silica Gel. A list of common mobile phases in order of increasing eluting power. Applications and Summary TLC has a number of practical applications in the laboratory.

TLC may be used to identify unknown compounds and unknown components of mixtures via comparison with standards. thin layer chromatography method


TLC is commonly used to monitor the course thin layer chromatography method a chemical reaction, and to assess the purity of the product through the comparison of relative amounts of reactants, products, and by-products on successive chromatograms over time.

TLC can also be used to determine if a substance purified by other methods such as recrystallization or distillation still contains a significant amount of impurity. When similar compounds cannot be resolved with TLC, they can be further separated based on another thin layer chromatography method property, known as a two-dimensional separation.