Related Experiment Video
Updated: Aug 20, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Effect of temperature and flow-rate on analysis of basic compounds in high-performance liquid chromatography using a
1Faculty of Applied Sciences, University of the West of England, Bristol, UK. David.Mccalley@uwe.ac.uk
Abstract:
The peak shape and retention of some basic probes together with a neutral reference compound were investigated as a function of temperature and flow-rate using a reversed-phase HPLC column at both pH 3.0 and pH 7.0. The retention of bases often showed an anomalous increase with temperature; retention mechanisms are complex as shown by studies of the effect of buffer cation concentration on retention. Considerable improvements in column efficiency for bases may result from operation at elevated temperature. Improvements did not seem attributable either to incidental changes in the retention factor, or (in this particular study where low sample masses were utilised) to the influence of sample load. The optimum flow-rate for highest efficiency is generally lower for basic compounds than neutrals, and due to the steepness of the Van Deemter curves obtained, high flow-rates appear to be particularly detrimental in the chromatography of basic compounds.
Related Concept Videos
Principles Of Column Chromatography
Column Efficiency: Rate Theory
During elution, a solute molecule experiences numerous transitions between stationary and mobile phases, exhibiting irregular residence times in...
Gas Chromatography: Types of Columns and Stationary Phases
For an analyte to remain on the column for a sufficient amount of time, it must exhibit some level of compatibility (or...
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
High-Performance Liquid Chromatography: Elution Process
High-Performance Liquid Chromatography: Instrumentation

