Related Experiment Video
Updated: Jun 9, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Interpreting the difference between conventional and bi-directional plate-height measurements in liquid
1Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250, USA.
Abstract:
An experimental and theoretical study was conducted of the column characterization technique in which plate heights determined using the conventional pulse-response method are compared with those determined using a bi-directional method where an eluite sample is introduced into one end of a chromatographic column and elution occurs at the same end after the flow direction is reversed inside the column. Experiments are presented for a micropellicular HPLC column before and after its performance has been degraded by repeated sample injections, for a low-pressure column containing nonporous glass particles, and for an HPLC column containing particles with 300 A pores. The results obtained are interpreted in terms of several different theories which apply in various Fourier number ranges. It was shown that the transcolumn contribution to convective dispersion in a chromatographic column is largely responsible for the difference observed between conventional and bi-directional plate-height measurements and that a collocation method can be employed to develop a useful analytical expression for this contribution.
Related Concept Videos
Column Efficiency: Plate Theory
A higher number of theoretical plates signifies better column efficiency and improved separation capabilities. Plate height affects bandwidth and separation quality; it is inversely...
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
Chromatographic Methods: Terminology
High-Performance Liquid Chromatography: Instrumentation
Chromatographic Resolution
The effectiveness of separation can be evaluated by determining the level of separation between two neighboring peaks in a chromatogram, which represents the individual components of a sample.
In chromatography,...
High-Performance Liquid Chromatography: Elution Process

