Stationary Phase and Mobile Phase Selection for Liquid Chromatography
CEN Webinars: Stronger Bonds
Stationary Phase And Mobile Phase
Selection For Liquid Chromatography
Tuesday October 21st 2014
8:00 a.m. PDT / 11:00 a.m. EDT / 16:00 BST
SPEAKERS
Speaker
Tony Edge,
R&D Principal
Thermo Fisher Scientific
 
Speaker
Dr. Shanhua Lin,
Program Manager
Thermo Fisher Scientific
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MODERATOR
Moderator
Britt Erickson,
Senior Editor,
C&EN
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OVERVIEW

This presentation focuses on how to choose the appropriate mode of separation, the correct column and also highlights the importance of the correct mobile phase. This approach will be applied to a wide selection of compound types ranging from proteins, peptides, glycans to small pharmaceutical molecules and their metabolites. The presentation will also look at specific application area for monoclonal antibody analysis, namely: titer, aggregation, charge variant, and oxidation variant. Platform methods for biologics characterization will be discussed as well.

The presentation will begin with a discussion on the structure and physiochemical properties of an analyte and how properties such as log D and pKa can help in column and mobile phase selection to use this to start to design the separation mechanism. The size of the analyte molecules will also be discussed in relation to choosing the correct type of support for the stationary phase, and the effect of pore size, surface morphology on the column loadability and chromatographic performance will be discussed.

The method development process will be expanded upon to include buffer selection and also the choice between gradient and isocratic separations will be discussed. The use of scouting gradients will be discussed in detail and how a separation based on this initial approach can be further optimized in simple and quick steps.

The presentation will also look at specific application area for monoclonal antibody analysis, namely: titer, aggregation, charge variant, and oxidation variant. Platform methods for biologics characterization will be discussed.

KEY LEARNING OBJECTIVES:

- How to choose the correct mode of chromatography

- Understanding of the fundamental retention mechanisms used in HPLC

- Understanding the workflow process for mAb analysis

- The effect that different solvents can have on the retention and separation of compounds

WHO SHOULD ATTEND
  • • Anybody who uses HPLC and wants to learn more about developing methods
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