Gradient

„The “secrets” of the gradient – Selection, Optimization, Troubleshooting“

 Target audience

Advanced HPLC-users in development departments, minimum experience 2 years

Training objectives

To gain certainty and ease of handling mobile phase and gradients

Training overview

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

Training content

  • 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