Chromatography Forum

Hi all,

After 2 years working in a university lab as the HPLC technician, I want to start testing the viability of my columns (mainly C18). Please can anyone suggest an appropriate test mix and conditions?

Thanks

I would make up or purchase the mixture as shown in the test chromatogram supplied with the column, and use those conditions. Do this when column is new, and store to compare with chromatograms as the column "ages".

That is good advice. Speaking as a column manufacturer, that is one reason why we give you the test conditions.

Thanks for your quick replies.

Part of my problem is that many of the columns do not have these test mixtures with them. Is there a generic mixture I can use which will show me whether the column is stii viable?

Still viable, I meant.

Alltech and Supelco (and probably other vendors) sell the HPLC test mixtures. You can choose the one most appropriate for your samples or column uses. Use the same mixture and test the column when brand new then you can compare a questionable column to the original conditions.

The advice above is reasonable-that is to use the original manufacturer test conditions and compounds to assess the column, or to use the Alltech or Supelco mixtures. These tests will show you if there are serious problems with any of your columns. However, many of the test conditions/compounds in such mixtures will not show up problems of column ageing and deterioration. Some manufacturer tests are specifically designed to show the column up in a good light. Thus the manufacturer test mix may still run o.k. but the column may no longer work for your analysis.

If you have the time, and you are not just trying to show up gross column problems, I would suggest you design your own test around the sorts of analysis that you are performing in your own lab-then you will have a very clear idea of whether the column is still good for a particular analysis.

Thanks for your help everybody,
I think I'll buy a test mixture and go for that, it should show all I need for undergraduate teaching /project purposes.