This technical article looks at diagnosing problems with our HPLC separations.
If we look at change in the chromatogram and concentrate on what happens to the retention time, tR, and the column dead time, t0, there are only four possible combinations: both t0 and tR change together, only tR changes, only t0 changes (highly unlikely), and neither variable changes (no change = no problem). Only the first two are of interest. First, let’s consider what could be the matter if both retention and the column dead-time change together. It has to be either a flow-rate-related change or a change of the column size.
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