The ability to identify and correct problems with HPLC or UHPLC separations is an important skill for practising chromatographers. In addition to the mechanical or electronic problems that can compromise the performance of instrument modules, changes in the chemical or physical characteristics of the column and/or unintentional changes in mobile phase chemistry can alter the quality of the separation.
This one-day course is designed to equip analytical scientists with the knowledge and tools required to correctly diagnose mechanical or chemical problems in HPLC or UHPLC separations. The course includes details of simple tests that can be performed to help identify the cause of the problem as well as simple precautions that can be taken to prevent faults occurring in the first place.
Course Programme
Chromatographic Theory and Troubleshooting Principles
This one-day course is designed to equip analytical scientists with the knowledge and tools required to correctly diagnose mechanical or chemical problems in HPLC or UHPLC separations. The course includes details of simple tests that can be performed to help identify the cause of the problem as well as simple precautions that can be taken to prevent faults occurring in the first place.
- Review of HPLC theory as it applies to troubleshooting and instrument maintenance
- Operating principles of each module in a HPLC/UHPLC system
- Performance qualification (PQ) is so important
- Techniques for systematic problem solving and instrument maintenance
- Preventing common hardware problems and method failures
Course Programme
Chromatographic Theory and Troubleshooting Principles
- Causes of adsorptive interactions in HPLC
- Chromatographic behaviour of acids and bases
- Revision of chromatographic theory
- Record-keeping
- Tips for effective troubleshooting
- Using information from system suitability tests
- Tubing and extra-column dispersion
- Different pump types
- Common problems with HPLC pumps
- Flow rate and leak tests
- Common detector types and characteristics
- Noise and drift tests
- Mobile phase absorbance in UV detection
- MS and evaporative detector-compatible mobile phases
- Effect of flow-cell volume
- Selecting appropriate data system settings
- Mobile phase filtration – when and how?
- Deaeration
- Buffer selection
- Mixing solvents – miscibility and other considerations
- Mobile phase stability
- Gradient mixing and delay volume
- Gradient performance qualification tests
- Autosampler types
- Common autosampler problems
- Causes of poor precision
- Curing carryover
- Column temperature effects
- Typical problems/symptoms
- Sample matrix effects
- Injection solvent composition and volume injected
- Column ageing
- Causes of peak asymmetry and retention time shifts
- Causes of high back pressure; sample filtration
- Sample stability and analyte adsorption
- Tests to troubleshoot problems with separation chemistry
- Peak integration and smoothing
- Advantages of second derivative plots
- Common integration errors
