Advances in Chromatography: Volume 59

This chapter discusses the thermodynamics and kinetics underlying hydrophobic interaction chromatography of proteins, the use of a kinetic model in predictive modeling of evaporation processes, building and employing QSRR models in cyclodextrin-modified HPLC, and reviews chemometric methods commonly paired with comprehensive 2D separations and key instrumental and preprocessing considerations.

Format: Hardback
Length: 200 pages
Publication date: 30 December 2022
Publisher: Taylor & Francis Ltd

Here is the rephrased text:

Hydrophobic interaction chromatography (HIC) is a powerful technique used to separate and purify proteins based on their interactions with hydrophobic surfaces. It relies on the principles of thermodynamics and kinetics to understand and manipulate the behavior of proteins in these environments.

In HIC, proteins are first immobilized on a hydrophobic solid support, such as silica or carbon, which allows them to bind to the surface and form stable complexes. The mobile phase, typically a hydrophobic solvent, is then passed through the column, causing the proteins to migrate based on their hydrophobic interactions.

One of the key advantages of HIC is its ability to separate proteins based on their hydrophobic properties, even when they share similar chemical compositions. This is because the hydrophobic interactions between the proteins and the support are stronger than other non-polar interactions, such as hydrogen bonding or ionic interactions.

A kinetic model can be used in the predictive modeling of evaporation processes, which eliminates the need to know the composition and identity of the chemical constituents in the sample. The model describes the rate of evaporation as a function of temperature, pressure, and the concentration of the evaporating species. By using this model, it is possible to predict the evaporation rate of a mixture without the need for extensive experimental measurements.

Building and employing QSRR (Quadratic Separation Range) models in cyclodextrin-modified high-performance liquid chromatography (HPLC) is another area of research in HIC. QSRR models are used to predict the retention behavior of proteins in HPLC, based on their amino acid sequence. These models can be used to optimize the separation conditions and improve the resolution of protein mixtures.

Chemoometric methods are commonly paired with comprehensive 2D separations in HIC. These methods involve the use of analytical techniques, such as mass spectrometry or fluorescence spectroscopy, to identify and quantify the proteins in the separated samples. By combining these methods with HIC, it is possible to obtain more detailed information about the protein composition and structure of the samples.

Key instrumental and preprocessing considerations must be taken into account when performing HIC. The choice of hydrophobic solid support, mobile phase, and separation conditions can have a significant impact on the separation efficiency and resolution of the protein mixtures. Additionally, proper sample preparation and handling are essential to ensure accurate and reproducible results.

In conclusion, HIC is a powerful technique used to separate and purify proteins based on their hydrophobic interactions. It relies on the principles of thermodynamics and kinetics to understand and manipulate the behavior of proteins in these environments. By using kinetic models, QSRR models, and chemoometric methods, it is possible to obtain more detailed information about the protein composition and structure of samples. Proper instrumental and preprocessing considerations must be taken into account to ensure accurate and reproducible results.

Weight: 540g
Dimension: 234 x 156 (mm)
ISBN-13: 9781032360270