100% Methanol in Size exclusion HPLC column Yarra sec 3000

Discussions about gel permeation chromatography / gel filtration chromatography / size exclusion chromatography

5 posts Page 1 of 1
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.
Gerhard Kratz, Kratz_Gerhard@web.de
Gerhard Kratz wrote:
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.
Gerhard Kratz, Kratz_Gerhard@web.de
Thanks for your suggestion. I am running 10% ACN now at .25 ml/min, I will report result here once I am done. Thanks !
5 posts Page 1 of 1

Who is online

In total there is 1 user online :: 0 registered, 0 hidden and 1 guest (based on users active over the past 5 minutes)
Most users ever online was 1117 on Mon Jan 31, 2022 2:50 pm

Users browsing this forum: No registered users and 1 guest

Latest Blog Posts from Separation Science

Separation Science offers free learning from the experts covering methods, applications, webinars, eSeminars, videos, tutorials for users of liquid chromatography, gas chromatography, mass spectrometry, sample preparation and related analytical techniques.

Subscribe to our eNewsletter with daily, weekly or monthly updates: Food & Beverage, Environmental, (Bio)Pharmaceutical, Bioclinical, Liquid Chromatography, Gas Chromatography and Mass Spectrometry.

Liquid Chromatography

Gas Chromatography

Mass Spectrometry