Related Experiment Video
Updated: Aug 15, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Sub- and supercritical chiral separation of racemic compounds on columns with stationary phases having different
Hiroko F Kasai1, Masayoshi Tsubuki, Sohichiro Matsuo
1Faculty of Pharmaceutical Sciences, Hoshi University, Tokyo, Japan. kasai@hoshi.ac.jp
Abstract:
Separation of the enantiomers of each of three different racemates, neutral rac-alpha-tetralol, acidic rac-2-phenylpropionic acid, and basic rac-1-phenylethylamine, using subcritical and supercritical fluid chromatography with two different chiral stationary phases, heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin (Sumichiral OA-7500 column) and tris(3,5-dimethylphenylcarbamate) of amylose (Chiralpak AD-H column), was compared. The elution order of the enantiomers of the three racemates was determined, and the effects of the type of alcohol modifier, column oven temperature, mobile phase composition, flow rate, and pressure were examined. The most appropriate column oven temperature depended on both the type of alcohol modifier and the compound analyzed. Lower alcohol content improved the peak separation of both rac-alpha-tetralol on the Sumichiral OA-7500 column and rac-1-phenylethylamine on the Chiralpak AD-H column, while the same phenomenon was not observed with either rac-alpha-tetralol or rac-2-phenylpropionic acid on the Chiralpak AD-H column. Decreasing outlet pressure improved the peak separation obtained with rac-2-phenylpropionic acid, but had little effect on either rac-alpha-tetralol or rac-1-phenylethylamine.
Related Concept Videos
Supercritical Fluid Chromatography
SFC utilizes a supercritical fluid mobile phase,...
Racemic Mixtures and the Resolution of Enantiomers
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...
Chromatography: Introduction
The phase in which the compounds linger or on which the compounds adsorb is called the stationary phase, whereas the mobile phase is the solvent that carries the solutes to be analyzed. In traditional column chromatography, the mixture flows through the stationary phase, and the compounds partition between the stationary and mobile phases...
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...
