A non-destructive test to assess the axial heterogeneity of in situ modified monoliths for HPLC

摘要

This paper describes a non-destructive a€?peak parkinga€? protocol in order to assess the axial heterogeneity of an in situ modified monolithic column for high performance liquid chromatography; a a€?gradient stationary phasea€? was designed whereby the ligand density decreases along the length of the rod in the a€?forward flowa€? configuration. The results of multi-location peak parking demonstrated a consistent increase in peak variance from the 1 cm position of the column to the 9 cm location. This increase in band broadening supported the theory of a decreasing ligand density along the length of this gradient column. This is consistent with efficiency measurements performed in both the forward and reverse flow directions, with an improved efficiency (15% increase in N ma?’1) in the reverse direction. These results are consistent with theoretical investigations into stationary phase gradients.