Chromatography Forum

Hi Guys

Accidentally 100% methanol was flow through Yarra SEC 3000 size exclusion column for long duration and even left on it over weekend. Now the separation performance is terrible. I thought sudden change in 100% organic solvent could have denatured the strongly adsorbed protein so I have flowed 6M Guanidine isothiocynate through it but it didn't restore the column performance. It was a newer column. Looks like it might have dehydrated due to methanol but I have been running aqueous solution for atleast 24hrs. Any suggestions.

The packing is destroyed due to swelling/shrinking of the packing material. Also silica with this special endcapping is effected. Please order a new column.

The packing is destroyed due to swelling/shrinking of the packing material. Also silica with this special endcapping is effected. Please order a new column.

Thanks for your response. This column can take upto 100% acetonitrile as per manufacturer instructions. Do you think i may have a chance to rehydrate it by running water slowly in reverse direction.

I'm quite sure that you will have a little void volume on the column head. If you are experienced chromatographer you may be able to repair it. Reversed Flow is a good idea, but only water to flush the column will be not enough. A little bit organic modifier is requested. Maybe 10% ACN can help.
Good luck.

Thanks for your suggestion. I am running 10% ACN now at .25 ml/min, I will report result here once I am done. Thanks !