This module is part of the Masterclass in Modern HPLC programme.
This is a rolling programme of six modules, which are run once every two months at six locations around the UK. Each module is complete in isolation, and there is no need to do the modules in any particular order. Simply select your preferred location, and in the following 12 months you will complete the course. |
A guide to the use of HPLC Solvents.
- Purity
- Viscosity
- Eluent Strength for normal phase and reversed phase
- Refractive Index
- Miscibility with other solvents
- Miscibility with water
- Sample solubility
- Buffer solubility
- Specific grades available
- UV transparency and UV cut-off
- Shelf life
- Specific Hazards - Toxicity, Flammability, Evaporation Rate
- Acid/Base/Dipole Selectivity Properties
- Compatibility with HPLC Instrumentation
Solvents to use as sample solvent
Solvents to use for sample extraction
Solvents to use for column cleaning
Solvents to use for column storage
A guide to the use of pH Buffers
- The importance of pH in HPLC
- When to use Ion Suppression and when to buffer
- pKa
- How a buffer works
- Selection of buffer concentration
- Which buffer to use at which pH
- Specifics about phosphates, acetates, borates, tris, amines, formates, citrates, glycines and ammonia
- UV cut-off
- Shelf life and storage
- Making up buffers - calculating concentration and adjusting pH
- Compatibilty with HPLC Instrumentation
Recommended Eluents
- Normal phase
- Reversed phase
- Ion exchange
- Ion chromatography
- GPC
- GFC
- LCMS
Making up an accurate mobile phase
Degassing
Effect of changing eluent composition
A guide to Ion Pair Reagents
- Anionic and Cationic
- Solubility
- Effect of alkyl chain length
- Use in enhancing detection
- Effect on equilibration times
- Use with gradient elution
- Concentration to use
- Troubleshooting
Eluent Optimisation
- Systematic solvent Optimisation using only 7 injections
- Optimisation of flow rate
Safety, Risk Assessment, Storage and Disposal. |