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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
A simple subcritical chromatographic test for an extended ODS high performance liquid chromatography column
Eric Lesellier1, Alain Tchapla
1Groupe de Chimie Analytique de Paris Sud, LETIAM IUT d'Orsay EA 3343, Plateau du Moulon, Orsay, France. eric.lesellier@iut-orsay.fr
Journal of Chromatography. A
|October 1, 2005
Summary
A new subcritical fluid chromatography (SFC) test using carotenoid pigments efficiently compares C18 stationary phase properties, including hydrophobicity and silanol activity. This method aids in selecting optimal High-Performance Liquid Chromatography (HPLC) columns.
Area of Science:
- Analytical Chemistry
- Chromatography
Background:
- Commercial C18 stationary phases exhibit variable properties impacting chromatographic performance.
- Standardized methods for comparing these properties are crucial for reproducible High-Performance Liquid Chromatography (HPLC) analyses.
Purpose of the Study:
- To introduce a novel subcritical fluid chromatography (SFC) test for comprehensive evaluation of C18 stationary phase characteristics.
- To establish a classification system for C18 columns based on their chromatographic behavior.
Main Methods:
- Development and validation of an SFC-based test utilizing carotenoid pigments as probes.
- Analysis of 129 commercial C18 columns to determine absolute hydrophobicity, silanol activity, and steric separation factor.
- Correlation of SFC test results with established HPLC and SFC methods (Tanaka, Engelhard, Sander, Wise).
Main Results:
- The SFC test provides a single-run assessment of key stationary phase properties.
- A classification diagram based on two separation factors effectively groups columns with similar chromatographic behaviors.
- Eight distinct column groups were identified, categorized by phase type (polymeric vs. monomeric), pore size, and endcapping.
Conclusions:
- The developed SFC test is a rapid and effective tool for comparing C18 stationary phases.
- The classification system aids researchers in selecting appropriate HPLC columns based on specific application needs.
- This method offers valuable insights into stationary phase characteristics, facilitating informed column choices.
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