Multi-Dimensional Liquid Chromatography: Principles, Practice, and Applications

A 2D-LC method is a powerful tool in analytical chemistry that can be used to separate, identify, and quantitate a wide range of compounds in complex samples. It offers several advantages over 1D-LC, including increased resolution, improved selectivity, and the ability to analyze more samples in less time. This article provides a comprehensive guide to the technique, covering its principles, methods, and applications. It also highlights the recent literature advances and practical guidance that can help researchers develop high-quality 2D-LC methods that meet their specific needs.

Format: Hardback
Length: 390 pages
Publication date: 30 December 2022
Publisher: Taylor & Francis Ltd

Here is the rephrased text:
Demonstrates the conditions under which a 2D-LC method should be considered as an alternative to a 1D-LC method.

Establishes a sound fundamental basis of the principles of the technique, followed by guidelines for method optimization.

Provides a single source for technical knowledge advances and practical guidance described in recent literature.

Assists with the initial decision to develop a 2D-LC method.

Guides the reader in developing a high-quality method that meets the needs of their application.

The development of 2D-LC methods has revolutionized the analysis of complex biological samples, offering increased resolution, sensitivity, and selectivity. In this comprehensive review, we will explore the principles, techniques, and applications of 2D-LC, highlighting its advantages over traditional 1D-LC methods.

Principles of 2D-LC:

2D-LC is a separation technique that utilizes two dimensions of separation, typically time and/or space, to achieve high resolution and selectivity. The separation is achieved by using a mobile phase, which is composed of a solvent and a stationary phase, and applying a suitable separation mechanism, such as size exclusion, ion exchange, or hydrophobic interaction.

One of the key advantages of 2D-LC is its ability to separate complex mixtures of proteins, peptides, and other biomolecules. By using two dimensions, the technique can resolve proteins that are closely related in size and charge, allowing for more accurate and comprehensive analysis of protein complexes. Additionally, 2D-LC can separate proteins based on their post-translational modifications, such as phosphorylation, glycosylation, and proteolysis, providing valuable insights into protein function and regulation.

Another advantage of 2D-LC is its ability to separate complex mixtures of lipids, such as phospholipids, triglycerides, and cholesterol. By using different mobile phases and stationary phases, 2D-LC can separate lipids based on their hydrophobicity, polarity, and size, providing insights into lipid metabolism and lipid-protein interactions.

Techniques of 2D-LC:

2D-LC can be performed using a variety of techniques, each with its own advantages and limitations. Some of the most common techniques include:

Size Exclusion Chromatography (SEC): SEC is a widely used 2D-LC technique that separates molecules based on their size. The mobile phase is composed of a solvent that has a larger molecular weight than the molecules being separated, and the stationary phase is a porous material that traps larger molecules. SEC is fast, efficient, and can be used to separate a wide range of molecules, including proteins, peptides, and lipids.

Ion Exchange Chromatography (IEC): IEC is a 2D-LC technique that separates molecules based on their ionic charge. The mobile phase is composed of a buffer solution that contains ions that are exchanged with the molecules being separated. The stationary phase is a material that binds the exchanged ions. IEC is useful for separating proteins that have different charge properties, such as acidic and basic proteins.

Hydrophobic Interaction Chromatography (HIC): HIC is a 2D-LC technique that separates molecules based on their hydrophobicity. The mobile phase is composed of a non-polar solvent that is immiscible with water, and the stationary phase is a hydrophobic material that traps hydrophobic molecules. HIC is useful for separating lipids that are not soluble in water, such as phospholipids and cholesterol.

Reverse Phase Chromatography (RPC): RPC is a 2D-LC technique that separates molecules based on their polarity. The mobile phase is composed of a polar solvent that is immiscible with water, and the stationary phase is a non-polar material that traps polar molecules. RPC is useful for separating proteins that have different polarity properties, such as hydrophobic and hydrophilic proteins.

Applications of 2D-LC:

2D-LC has a wide range of applications in biochemistry, biophysics, and medicine. Some of the most common applications include:

Protein Analysis: 2D-LC is used to analyze complex protein mixtures, such as those obtained from cell lysates or tissue samples. By separating proteins based on their size, charge, or post-translational modifications, 2D-LC can provide insights into protein function, protein-protein interactions, and protein turnover.

Lipid Analysis: 2D-LC is used to analyze complex lipid mixtures, such as those obtained from blood or tissue samples. By separating lipids based on their hydrophobicity, polarity, and size, 2D-LC can provide insights into lipid metabolism, lipid-protein interactions, and lipid-related diseases.

Drug Discovery: 2D-LC is used in drug discovery to identify and characterize potential drug candidates. By separating drug candidates based on their physicochemical properties, 2D-LC can provide insights into drug metabolism, drug-protein interactions, and drug efficacy.

Conclusion:

2D-LC is a powerful separation technique that has revolutionized the analysis of complex biological samples. By utilizing two dimensions of separation, 2D-LC can achieve high resolution, selectivity, and sensitivity, allowing for more accurate and comprehensive analysis of proteins, peptides, and lipids. The technique has a wide range of applications in biochemistry, biophysics, and medicine, and its development has opened up new avenues for research and discovery. As the field of 2D-LC continues to evolve, it is likely to play an increasingly important role in advancing our understanding of biological processes and developing new therapies for diseases.

Weight: 948g
Dimension: 181 x 262 x 26 (mm)
ISBN-13: 9780367547660