Separation Through chromatography

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SUMMARY

Chromatography is a separation technique utilized primarily for analyzing mixtures rather than extracting components. Thin Layer Chromatography (TLC) and paper chromatography are effective for visualizing substances but are not ideal for extraction. For purification purposes, column chromatography is recommended as it allows for the separation of substances for further use. Techniques such as using DEAE-cellulose plates with NaCl eluant can enhance the visualization of components, particularly in the purification of nucleoside triphosphates.

PREREQUISITES
  • Understanding of chromatography techniques, specifically Thin Layer Chromatography (TLC) and paper chromatography.
  • Familiarity with column chromatography for substance purification.
  • Knowledge of DEAE-cellulose and its application in chromatography.
  • Basic principles of solvent interactions and separation methods.
NEXT STEPS
  • Research the principles and applications of column chromatography.
  • Learn about the preparation and use of DEAE-cellulose plates in chromatography.
  • Explore the visualization techniques for chromatographic results, including UV illumination.
  • Investigate the extraction methods for substances post-chromatography.
USEFUL FOR

Researchers, chemists, and laboratory technicians involved in separation processes, particularly those focusing on purification techniques in organic chemistry.

Deadevil
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Dear all,
We all know that chromatography is a separation technique. In paper or TLC,the mixture is differentiated according to their relative affinities with the solvent on the paper or glass plate. How do we really separate the mixture. Kindly, help me how to extract the components of mixture from paper or TLC.
Thanks
 
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TLC is not typically used for extraction, rather to check what is present in the solution/how is the reaction going. If you want to separate substances for further use, use column chromatography.
 
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You mean Paper and TLC..?
 
Yes.

In case small amount is enough, you can do the chromatography (TLC or paper), cut the stain and extract the substance in small amount of solvent. But in general that's not what TLC and paper chromatography is for.
 
Borek said:
Yes.

In case small amount is enough, you can do the chromatography (TLC or paper), cut the stain and extract the substance in small amount of solvent. But in general that's not what TLC and paper chromatography is for.

I often used to use TLC to purify nucleoside triphosphates , (DEAE-cellulose with NaCl eluant) - it was pretty simple, only small quantities were needed and you very easily see the bands of the triphosphates and of impurities by UV illumination. Just came back to me that before chromatogrphing the sample, we prepared the DEAE-cellulose plates putting the bottom of the plate in water or dilute salt and this pushed some stuff out of the DEAE which you could see as a dark band that moved with the front, then we dried them. Don't knows if many people would use it nowadays though.
 
I agree with Borek. Column chromatography is the way forward for typical purification processes. Let me know if you need help with that. :)

Cheers
 

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