Separation Through chromatography

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Discussion Overview

The discussion revolves around the separation of mixtures using chromatography techniques, specifically paper chromatography and thin-layer chromatography (TLC). Participants explore the effectiveness of these methods for extracting components from mixtures and consider alternative techniques for purification.

Discussion Character

  • Exploratory
  • Technical explanation
  • Debate/contested

Main Points Raised

  • One participant inquires about the process of separating mixtures using chromatography and seeks guidance on extracting components from paper or TLC.
  • Another participant suggests that TLC is primarily used for monitoring reactions rather than for extraction, recommending column chromatography for separation intended for further use.
  • A participant confirms the focus on paper and TLC and mentions that small amounts can be extracted by cutting the stain and using a solvent, but emphasizes that this is not the primary purpose of these techniques.
  • A further contribution describes a personal experience using TLC for purifying nucleoside triphosphates, detailing the preparation of DEAE-cellulose plates and the visibility of bands under UV light, while expressing uncertainty about the current prevalence of this method.
  • Another participant agrees with the recommendation for column chromatography as a more suitable method for typical purification processes.

Areas of Agreement / Disagreement

Participants express differing views on the appropriateness of TLC and paper chromatography for extraction purposes, with some advocating for column chromatography as a more effective method. The discussion remains unresolved regarding the best approach for extracting components from mixtures.

Contextual Notes

Participants mention specific techniques and personal experiences, but there are limitations in terms of assumptions about the effectiveness of various chromatography methods and the context in which they are applied.

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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