Back to Basics 2: Mobile Phase Selection In HPLC & UHPLC
Mobile phase composition is a fundamental decision which has a major impact on the separation of analytes in your HPLC or UHPLC method. It has an effect on retention time of analytes, selectivity between these and the peak efficiency. Consideration should be taken not only to the type of solvents used, but also the concentration of these and whether an isocratic run or a gradient run is most suitable. Additives can also impact the resolution, retention and peak shape of sample analytes and so careful choice of buffer must also be made. We discuss all of these topics, looking at the most commonly used organic modifiers for reversed phase U/HPLC in both isocratic and gradient methods and how these can impact your separation and run times.
A number of topics will be covered during the seminar:
- Solvent options for U/HPLC
- Method development solvent considerations
- Isocratic and gradient chromatography
- Influence of buffer salt and pH on separations