UPLC for SDS/BME containing protein sample

I am currently trying to transfer an old HPLC method from another group that uses approximately 2% SDS and 5% BME.  The old method uses a Bakerbond C8 column, 5 µ, 300A, 4.6 ´ 100 mm (Baker Cat. #7105-01) and I would like to use a UPLC column instead.  The column does not have to match this one exactly as far as ligand, but the 300A pore size is a must as this is a big protein.Any suggestions that anyone can offer would be great.  I have an Acquity BEH300 C4 column that I am tempted to try this on but am afraid of destroying the column.

Answers

  • Hello,

    The BEH300 C4 column is the column where the most work has been done for the analysis of intact proteins. As far as the mobile phase, we have not done any work with SDS and BME. However, please take a look at the attached pdf on developing separations for intact proteins using the BEH300 C4 column. This poster was from HPLC 2010 last year.

    Good luck,

    --Doug

  • Hi Doug,

    Thank you for your reply.

    The mobile phase will be 0.1% TFA in water and ACN, but the sample will contain SDS and BME.  Is there any data for column deterioration under these conditions?

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