Can the theory of gradient liquid chromatography be useful in solving practical problems?

Area of Science:

• Analytical Chemistry
• Chromatography

Background:

• Gradient liquid chromatography (GLC) is a powerful separation technique.
• Theoretical understanding of GLC is crucial for effective method development.

Purpose of the Study:

• To review advances in GLC theory and their practical impacts.
• To compare theoretical models for retention in various chromatographic modes.
• To discuss applications of theory in gradient method development, optimization, and transfer.

Main Methods:

• Comparison of theoretical models for reversed-phase, normal-phase, and ion-exchange chromatography.
• Focus on two- or three-parameter equations for fitting experimental data.
• Discussion of instrumental factors and sample properties affecting separations.

Main Results:

• Identification of practically useful models for gradient elution.
• Analysis of applications for method development, optimization, and transfer.
• Exploration of challenges such as dwell volume and sample size, with a focus on large molecules.

Conclusions:

• Theoretical models significantly aid in understanding and improving gradient liquid chromatography.
• Practical application of these theories facilitates robust method development and transfer, particularly for complex separations like those involving large molecules.