Chromatography Problem: Setting a "Clen After Gradient

In summary, the speaker is seeking clarification on a step in their chromatography protocol that instructs them to set a "clen" after gradient value to 100% buffer B over 2 CV. They ask if anyone knows what this means and suggests checking the manual for definitions of "CV" and "clen". They also mention that the chemistry forum may be a better place to ask for help with column chromatography.
  • #1
mountain
53
0
hi guys!
as you know i am doing the chromatography stuff and am now having the problem with understanding the protocol. it says;

"set a clen after gradient value to 100% buffer B over 2 CV”? :bugeye:
does any of you know what it tries to tell me?

thanks!
 
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  • #2
Does your manual spell out what they mean by "CV" anywhere? I could guess a few things, but probably would guess wrong. What about "clen"...is that a typo or what is actually written (or maybe what's actually written is a typo...been known to happen).

All these column chromatography questions might get better responses in the chemistry forum, since organic chemists use them more frequently (and such methods are almost always taught in organic chemistry classes).
 
  • #3


Hi there,

The "clen after gradient" refers to the cleaning step after the gradient has been run. The protocol is asking you to set the machine to run 100% buffer B (the mobile phase) for 2 column volumes (CV) in order to clean out any remaining sample or impurities from the column. This step is important for maintaining the integrity of your column and ensuring accurate results in future runs. I hope this helps clarify the protocol for you. Good luck with your chromatography!
 

1. What is "Clen" in chromatography and why is it important to set it after the gradient?

"Clen" refers to the cleaning solvent in chromatography that is used to remove any residual contaminants or impurities from the system. It is important to set the "Clen after gradient" to ensure that the system is properly cleaned before the next sample is injected, avoiding any carryover effects and ensuring accurate results.

2. What is a gradient in chromatography and why is it necessary?

A gradient in chromatography refers to the gradual change in the composition of the mobile phase (solvent) used to elute the sample components from the stationary phase (column). This is necessary to separate complex mixtures and improve the resolution of the sample components.

3. How do you set the "Clen after gradient" in chromatography?

To set the "Clen after gradient" in chromatography, you need to first run the gradient program with the desired mobile phase composition. Once the gradient is complete, switch to the cleaning solvent (Clen) and run it through the system for a sufficient amount of time to clean the column and detector. Finally, switch back to the initial mobile phase composition for the next sample injection.

4. What are the consequences of not setting the "Clen after gradient" in chromatography?

If the "Clen after gradient" is not set in chromatography, there is a risk of carryover effects from previous samples, which can lead to contaminated results and affect the accuracy of the analysis. Additionally, not cleaning the system properly can result in clogged columns and degraded performance.

5. How often should the "Clen after gradient" be set in chromatography?

The frequency of setting the "Clen after gradient" in chromatography depends on the specific system and the samples being analyzed. Generally, it is recommended to set the "Clen" after each gradient run to ensure the system is properly cleaned and to avoid any carryover effects. However, if the samples being analyzed do not have any potential for contamination, the "Clen" may be set less frequently.

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