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- Posts: 33
- Joined: Fri Nov 17, 2006 4:50 am
- Location: Australia
It needs to be a very sensitive method (<<50ug/mL).
I have had issues in the past and had developed two separate HPLC methods (using SEC for the protein) and a C8 100A for the peptide.
I only had a C8 column on hand at the time.
But, I was wondering if I could analyse them both on a C4 column, possibly in the one method.
However, would I need a 300A pore size column because of the protein? Would this mean the peptide would take ages to elute? I thought a C4 column would be good because I wouldn't require as much acetonitrile to elute the protein, and hence it would not crash out.
I have also learnt in order to keep the peptide in solution (because of the attached lipids) to heat the auto-sampler and the oven compartment to 37C.
I was wondering if there was an ideal ~2um; 100x2.1; ??Angstroms; perhaps coreshell technology UPLC column available. A require a very sensitive column. I have attached a high sensitive flow cell to the Shimadzu DAD.
I would very much appreciate any suggestions. Protein and peptide analysis is relatively new to me.