Related Experiment Video
Updated: Jul 1, 2026

Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis
Published on: July 11, 2014
Combined effect of temperature and organic modifier concentration on the retention under single mode gradient
Adriani Pappa-Louisi1, Panos Nikitas, Christiana Zisi
1Laboratory of Physical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki, Greece. apappa@chem.auth.gr
Abstract:
The combined effect of temperature, T, and organic modifier concentration, phi, on the retention under gradient conditions in RPLC is studied by considering, both theoretically and experimentally gradients, of phi at constant T and gradients of T at constant phi. Two approaches are examined: in the first approach the prediction of the elution time of a sample solute is based on the isocratic/isothermal properties of this solute. The second approach is based on a direct fitting procedure of a proper retention model to 2-D isocratic/T-gradient or isothermal/phi-gradient retention data. These approaches were tested using alkylbenzes in eluting systems modified by ACN. We found that both approaches can give excellent predictions under certain prerequisites. However, the first approach exhibits the notable advantage that it can be used effectively to predict retention times under any kind of phi-gradients at constant T or T-gradients at constant phi. The second approach has the advantage that it is relatively simple but its applicability is very restricted since its predictions are satisfactory only if the gradients are of the same kind with those used in the fitting procedure and the conditions lie within those used for fitting.
More Related Videos
06:50An Experimental Protocol for Studying Mineral Effects on Organic Hydrothermal Transformations
Published on: August 8, 2018
14:42Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems
Published on: September 23, 2021
Related Concept Videos
Optimizing Chromatographic Separations
Band broadening refers to spreading solute bands as they travel through the column. This broadening can impact resolution. Plate height (H) represents the length required for one theoretical plate. A lower plate height corresponds to...
Analyte Adsorption and Distribution
High-Performance Liquid Chromatography: Elution Process
Effect of Temperature Change on Reaction Rate
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
Chromatographic Methods: Classification
Chromatographic techniques are typically named by...