HPLC Method Development
For the experienced chromatographer, this course provides a comprehensive, process-driven approach to the development of an optimal and robust HPLC method.
The course includes a detailed exploration of each critical parameter within a typical method, then builds this into step-by-step process for method development. We apply relevant theory to underpin a practical approach which maps out the optimal route to a commercial quality method. Importantly we look at building robustness into each stage of development using a Quality by Design approach, highlighting many common problems and how we develop these out.
● Setting method development objectives
● Sample preparation
● System choices
● Choosing a column and mobile phase
● Optimising strategies
● Quantitation and system characterisation
- Design specification
- What is QBD?
- Building in robustness
- Retention factor
- Influencing sample solubility
- Choosing a diluent
- Sample polarity
- Organic modifiers
- pH control
- Physical dimensions
- Particle type, sizes and UHPLC
- Chemically modified silica
- Reversed phase ligands
- Hydrophobic subtraction model classification
- Isocratic or gradient
- Optimising k’
- Effect of each gradient parameter
Building a Practical Process
- Sample preparation
- Screening for alpha
- Chromatographic optimisation
- Linearity, range and sensitivity
This course is designed for the more experienced chromatographer with a good working knowledge of HPLC separations. Completion of Understanding HPLC and HPLC Troubleshooting training courses would be advantageous.
Delegates should have a good knowledge of chromatography and experience as HPLC users. Some experience in method development is recommended. A good grounding in chemistry is also beneficial.
Analytical Method Validation