Column capacity

[Gabriella]

I am trying to determine the maximum amount of protein (by mass) I can load onto a column and still get decent seperation. Is there a simple way to do a rough calculation? The column we are currently using is a 5 cm, 1 mm ID, column with 3.5um particals from agilent (zorbax 300SB-C18 column). I don't know for sure where I will have peaks since we are digesting proteins for MS analysis.

Gabriella

[tom jupille]

I am trying to determine the maximum amount of protein (by mass) I can load onto a column and still get decent seperation. Is there a simple way to do a rough calculation? Gabriella

No simple way to estimate because it will depend on how much resolution you need and how well separated your peaks are. If this were my problem, I would do a "quick and dirty" loading study (start low and increase by successive 3x multiples) with a "typical" sample.