Size Exclusion on the UPLC

We've been using a Size Exclusion column on our HPLC which runs with isocratic 50% ACN with 0.1% TFA to analyze wheat proteins.  The Waters Size Exclusion columns for the UPLC use a maximum of 20% organic.  We're concerned that our proteins will come out of solution with only 20% organic but we would really like to try the UPLC.  Given the cost of the columns we are reluctant to order one just to possibly have it fail.  Has anyone transferred a HPLC-SE method to the UPLC?  If so, what are your conditions?  Any help would be appreciated.

Answers

  • The Care and Use Manual for the Waters ACQUITY BEH200 SEC column recommends <20% methanol in the mobile phase only because the solubility of some proteins is low at high organic concentrations.  If your specific HPLC method is already known to be successful using 50% ACN with 0.1% TFA, then you should have no problem trying out your method on the Waters UPLC column.

  • Do you know what would be the highest concentration of organic solvent (acetonitrile or methanol) you could use with those columns?

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