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Sample doesnt stick to C18 column

Posted: Tue Nov 12, 2013 1:42 am
by tifaniin
Hi,
I am trying to synthesize and purify Shikimate-3-phosphate (S3P) using a C18 column. The reaction that I use for the synthesis of S3P contains shikimate, ATP, KCl, MgCl2 and Tris pH 7.0. It might due to the presence of the buffer components, but the sample does not stick to the column and comes straight off. I am using 0.1% TFA as mobile phase. Could anyone please advice me as to how I could make my sample stick to the C18 column? Thank you

Re: Sample doesnt stick toe. C18 column

Posted: Tue Nov 12, 2013 1:54 am
by tom jupille
Possibly addition of an ion-pairing reagent (e.g., tetrabutylammonium) would work, but my guess is that separation would be much easier using HILIC .

Re: Sample doesnt stick to C18 column

Posted: Thu Nov 14, 2013 11:26 pm
by Vlad Orlovsky

Re: Sample doesnt stick to C18 column

Posted: Wed Nov 20, 2013 2:44 pm
by danko
Using ion pair in purification context is not my prefered solution.
I would much rather go for Ion Exchenge mode.

Best Regards

Re: Sample doesnt stick to C18 column

Posted: Mon Dec 23, 2013 7:15 am
by Arne
It's likely too polar for C18 RP separation. Ion exchange will be the way to go imo.