Two-dimensional liquid chromatography with reversed phase in both dimensions: A review

Area of Science:

• Analytical Chemistry
• Chromatography

Background:

• Two-dimensional liquid chromatography (2D-LC), especially comprehensive (LC×LC), offers superior peak capacity, resolution, and selectivity over 1D-LC.
• Orthogonal separation mechanisms are typically preferred in 2D-LC, but using similar mechanisms, like reversed-phase (RP) in both dimensions (RPLC×RPLC), is gaining traction.
• RPLC×RPLC addresses challenges like mobile phase incompatibility and poor column efficiency, making it promising for complex sample analysis.

Approach:

• This review focuses on the physical and practical aspects of method development for RPLC×RPLC.
• It provides a comprehensive overview of RPLC×RPLC applications in various scientific fields.
• The review highlights the benefits of using reversed-phase columns in both dimensions for enhanced separations.

Key Points:

• RPLC×RPLC offers significant advantages for separating complex mixtures.
• Method development considerations for RPLC×RPLC are discussed.
• The technique is applicable across diverse fields, including food, natural products, environmental analysis, proteomics, lipidomics, and metabolomics.

Conclusions:

• RPLC×RPLC is a powerful technique for complex sample analysis, overcoming limitations of traditional 1D-LC.
• The review emphasizes the practical considerations and broad applicability of RPLC×RPLC.
• This approach provides enhanced separation capabilities for various analytical challenges.