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Reversed-phase pH gradient thin-layer chromatography of biologically active substances with controlled developing
Aneta Hałka-Grysińska1, Adrian Szczyrba1, Tadeusz H Dzido1
1Department of Physical Chemistry, Medical University of Lublin, Chodźki 4a, 20-093 Lublin, Poland.
Journal of Chromatography. A
|May 26, 2021
Summary
This study introduces stepwise pH gradient thin-layer chromatography for separating biologically active substances. Optimizing mobile phase pH is key to enhancing separation efficiency and spot characteristics in chromatography.
Area of Science:
- Analytical Chemistry
- Chromatography Science
Background:
- Thin-layer chromatography (TLC) is a common separation technique.
- Optimizing mobile phase composition is crucial for effective separation.
Purpose of the Study:
- To present stepwise pH gradient thin-layer chromatograms for biologically active substances.
- To investigate the impact of mobile phase pH on chromatographic separation.
Main Methods:
- Stepwise pH gradient thin-layer chromatography with controlled solvent velocity.
- Reversed-phase high-performance thin-layer chromatography (RP-HPLC).
Main Results:
- pH gradients significantly influence spot retardation, selectivity, and shape.
- Mobile phase pH is a critical factor for optimizing separations.
- Satisfactory reproducibility of gradient retardation factor values was achieved.
Conclusions:
- Stepwise pH gradient TLC is a novel and effective method for separating biologically active compounds.
- This technique offers enhanced control and optimization possibilities in chromatography.
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