When we encounter mixtures of substances while working as biologists, our first thought is always, "What are the components in this mixture? Chemical assays are a possibility for determining the presence of particular chemicals. But doesn't that seem painful?
The good news is that thin-layer chromatography (TLC) makes it simple to separate components from mixtures with the help of TLC plates and quickly identify the chemicals in a sample by employing developing agents and Rf values.
The following are some special applications of thin-layer chromatography in biology:
Detecting a Particular Compound in a Mixture
Biologists frequently extract substances from microbial, animal, or plant sources, typically resulting in a combination of substances. After extraction, your main goal would be to check for the existence of your target component. By running the standard and extract simultaneously and comparing their Rf values, you can use TLC to search for your compound if you have a pure form of the compound.
Two Compounds from a Different Origin Are the Same
Likewise, you can compare samples from several sources to see if the two substances are identical. The key would be identical Rf values.
Number of Compounds in a Mixture
A combination of compounds can be separated using TLC, and the number of compounds contained in the mixture can also be counted. To estimate the number of chemicals in the mixture, count the dots that formed on the TLC plate.
Solvent for Column Chromatography to Separate Compounds
To help you choose the solvent to utilize for column chromatographic separation of your compound of interest, you can use TLC to evaluate various solvent systems for your compound. To purify the compound using column chromatography, the solvent system that provides the best resolution during TLC can also be used.
Monitoring Reactions
By performing TLC on the reaction sample at specific intervals, you can keep an eye on the direction and rate of the reaction. For instance, you can gather reaction mixtures at various time intervals while executing compound biotransformation and use TLC to track the transformation of the reactants into products.
Checking Column Chromatographic Separations
By getting samples of the purified chemicals and using TLC, you can evaluate the effectiveness of column chromatographic separations. For instance, you can use TLC to assess several elutions' purity when using column chromatography to purify a pigment from an extract.
Try High-Performance Thin Layer Chromatography to Enhance Your TLC Results
You can get better resolution due to automation. A machine that automatically applies samples is used in HPTLC, a TLC variation. This strictly applies an identical amount of the sample to every experiment while ensuring correct sample application. Given these applications, TLC is a handy analytical method in biology. So, start TLC by using TLC plates and reduce your efforts.
