The determination of the plate count for large molecules under reversed-phase gradient conditions

The true performance of HPLC columns in the gradient separation of macromolecules can be assessed by measuring the true plate count, if we know the retention factor of the analyte at the point of elution. We are demonstrating in this short communication how this can accomplished in a straightforward fashion. The procedure used here is a significant simplification over previous approaches, and enables the chromatographer to do so without complex algebra or additional experiments.