„The “secrets” of the gradient – Selection, Optimization, Troubleshooting“
Advanced HPLC-users in development departments, minimum experience 2 years
To gain certainty and ease of handling mobile phase and gradients
First we will discuss the criteria of the „right“ mobile phase for a given separation problem. We will also talk about alternatives, the role of modifiers and the pH-value. The emphasis of the training adresses gradient elution. The participants will learn light-to-implement optimization tools as well as pitfalls and will hear about gradient myths and strange gradient stories… With the help of a numerous real examples one can understand the provided to-do-lists that give hints on how to create generic gradients depended on the number of compounds to be separated, the matrix and the analytical target
- Methanol or acetonitrile? And: Are organic solvents interchangeable?
- I need a pH-value of 3: What should I use – phosphoric, formic or acetic acid?
- Aspects of optimization – customary/unusual
- A lot of small peaks, related and polar compounds: Stepper gradient and low flow rate or vice versa?
- Some particularities of the gradient
- What does optimization of the gradient mean (resolution, peak capacity, low LOQ)?
- Myths, „unexpected“ outcomes, examples
- The problem of method transfer (HPLC/UHPLC, High-/Low-Pressure-Gradient, different vendors)
- Drifting baseline, “humps”, low RSD – how to proceed
- „Do ́s and don’t ́s“, list of recommendations