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[HPLC] HPLC Column Regeneration -
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Column Regeneration
HPLC에서 컬럼을 오래 사용하다보면 컬럼의 노화가 진행되어 분리능이 떨어지고 효율이 떨어집니다.
이러한 컬럼을 Regeneration 하는 방법에 대한 몇가지 설명입니다.
Particulate matter caught on the inlet frit
This can sometimes be removed by reversing the column and flushing to waste. An 0.5-µm porosity in-line filter will help to prevent this kind of problem, as will filtering or centrifuging your samples.
This can sometimes be removed by reversing the column and flushing to waste. An 0.5-µm porosity in-line filter will help to prevent this kind of problem, as will filtering or centrifuging your samples.
Loss of bonded phase
Some of this happens naturally over time and sometimes is accelerated by mobile phase pH outside the normal 2 < pH < 8 recommended operating conditions. If bonded phase loss occurs, there is no way to fix it.
Some of this happens naturally over time and sometimes is accelerated by mobile phase pH outside the normal 2 < pH < 8 recommended operating conditions. If bonded phase loss occurs, there is no way to fix it.
Build-up of insoluble materials on the column
If these are indeed insoluble, they cannot be washed off. If they are just poorly soluble in the mobile phase, such as fats, waxes, etc, sometimes they can be washed off by reversing the column and after flushing with ACN or MeOH, flushing with 50 mL methylene chloride, then back through ACN or MeOH to the mobile phase. If you suspect the nature of the contaminant, sometimes a particular solvent can be chosen that will be most effective at removing it. For example, proteins might be removed by adding a chaeotrope, such as guanidine, to the mobile phase. I suspect that recommendations for DMSO fall in this category.
If these are indeed insoluble, they cannot be washed off. If they are just poorly soluble in the mobile phase, such as fats, waxes, etc, sometimes they can be washed off by reversing the column and after flushing with ACN or MeOH, flushing with 50 mL methylene chloride, then back through ACN or MeOH to the mobile phase. If you suspect the nature of the contaminant, sometimes a particular solvent can be chosen that will be most effective at removing it. For example, proteins might be removed by adding a chaeotrope, such as guanidine, to the mobile phase. I suspect that recommendations for DMSO fall in this category.
(이하생략)
원문 출처 : http://www.sepscience.com/Techniques/LC/Articles/264-/HPLC-Solutions-100-Column-Regeneration