Thin Layer Chromatography | Principle
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- Опубликовано: 19 сен 2018
- This Lecture explains about Thin Layer Chromatography technique in Hindi.
Thin layer chromatography is a separation technique, which separates out individual compounds from the mixture depending upon the polarity of each compound.
Procedure: TLC Can be performed on the glass slide by applying a thin layer of silica gel. Once the plate is ready, draw a line using pencil, approx 1 cm from the base of the plate. Apply a small spot of the sample to be analyzed on the centre of the line, then place the TLC plate vertically in the chamber. The solvent system travels up a silica plate by capillary action and passes over the sample that spotted onto the plate. As the solvent travels up, it moves the compounds present in the sample at different rate depending on their polarity.
Rf valve is the ratio of distance traveled by the compound to the distance travelled by the solvent and ranges between 0 to 1. Every compound has a significant RF valve, allowing you to separate and identify them.
If the sample that is run on TLC plate is coloured, it can be visualized directly after the run. But often the sample is colourless. Therefore you will need to choose an appropriate technique to visualise the spots after TLC run. Some commonly used techniques are:
UV visualisation
Acid Treatment
Iodine Vapor Treatment
Vanillin stain
Ninhydrin treatment
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Is there any method to apply the sample without increasing the concentration? I mean, how to apply accurate amount of sample in the TLC plate?
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Different Compounds have different Polarity, You Can adjust the mobile phases According to the Polarity of Compounds, For Eg Meoh/ Mdc. EA/Pet ether
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