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High-performance chromatofocusing using linear and concave pH gradients formed with simple buffer mixtures. I. Effect
1Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County, Baltimore 21250, USA.
Journal of Chromatography. A
|September 8, 2000
Summary
This study develops a theoretical and experimental basis for selecting buffer compositions in chromatofocusing. It explains how buffering capacities influence pH gradient shapes for optimal separation.
Area of Science:
- Analytical Chemistry
- Separation Science
Background:
- Chromatofocusing is a powerful separation technique.
- Controlling pH gradient shape is crucial for optimizing separations.
- Understanding the factors influencing gradient formation is essential.
Purpose of the Study:
- To determine factors governing pH gradient shape in chromatofocusing.
- To establish the relationship between gradient shape and buffering capacities.
- To provide a basis for selecting buffer compositions in chromatofocusing.
Main Methods:
- Numerical calculations and simplified analytical relations based on local equilibrium theory.
- Investigation of buffering capacities in adsorbed and liquid phases.
- Experimental verification using high-performance chromatography columns.
Main Results:
- Identified key factors influencing pH gradient shape (linear or concave).
- Established a quantitative relation between gradient shape and buffering capacities.
- Experimental results validated the theoretical models.
Conclusions:
- The developed theoretical and experimental framework aids in selecting optimal buffer compositions for chromatofocusing.
- This work enhances the understanding and application of pH gradient control in separations.