The resolution is determining the ability to separate two analytes. It is a measure of how well two elution peaks can be differentiated in chromatographic separation.
The resolution, capacity factor, retention factor, column efficiency, and selectivity are the basic chromatography parameters. Resolution is determined by three important parameters such as efficiency, selectivity, and retention.
In this case, there is no specific parameter to apply, since each case is a special case in chromatography. Because in gas chromatography (GC) you can change with the column, flow rate and temperature gradient, the concentration of the mobile phase is always the same. As you know, liquid chromatography consists of separations according to the chemical interactions between the analytes and the stationary phase (column). Principally, the solute partitions amongst the moving phase and stationary phase. Each component has a different structure, pKa value, solubility, polarity, and pH, hence their different affinity toward the stationary phase they have different retention time. Consequently, you have to play with many different parameters to improve resolution in chromatography.
Various parameters are mentioned here, which you can play within HPLC to improve the resolution.
- The pH of the compound and mobile phase
- Column dimension, particle size, porosity, shape, and carbon loading
- Working and column temperature
- The flow rate of the system
- Ionic strength
- Organic concentrations of mobile phase
- Applied Method (Isocratic or gradient)
- The shape and speed of the gradient method
If you know that there is a difference in the structure between the molecules you want to separate, you can change the conditions accordingly to get better separation. If you do not have the compound identity, then you have to rely on your experience to make the appropriate changes for a better solution.
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