Theories to support method development in comprehensive two-dimensional liquid chromatography--a review.

Area of Science:

• Analytical Chemistry
• Chromatography Science

Background:

• One-dimensional liquid chromatography (LC) struggles with complex sample matrices.
• On-line comprehensive two-dimensional liquid chromatography (LC×LC) offers superior resolution for challenging samples.
• Optimization is key to realizing the full potential of LC×LC techniques.

Purpose of the Study:

• To critically review state-of-the-art strategies for optimizing on-line LC×LC.
• To clarify which aspects of LC×LC method development are currently optimizable.
• To guide practitioners in selecting appropriate chromatographic parameters.

Main Methods:

• Review of existing literature on LC×LC optimization strategies.
• Analysis of factors influencing resolving power, analysis time, and dilution.
• Identification of current limitations in predictive modeling for retention and orthogonality.

Main Results:

• Significant advancements have been made in optimizing efficiency-related parameters in LC×LC.
• Challenges remain in developing predictive models for retention factors and maximizing orthogonality.
• Current optimization often relies on post-analysis examination of chromatograms for orthogonality.

Conclusions:

• On-line LC×LC optimization strategies have progressed significantly.
• Further research is needed in predictive modeling of retention and orthogonality for enhanced LC×LC method development.
• A clear understanding of optimizable parameters is essential for effective LC×LC application.