Languages: English
Price (from): €1,400 / day
About the training
#Logical approach to method development #More efficient methods #Improved method robustness #Methods that are fit for purpose #Risk-based approach to method transfer #Reduced method development and transfer timelines
This course is intended for scientists undertaking HPLC method development, as well as those involved in transferring existing methods between laboratories. The course content assumes that attendees will have a basic understanding of HPLC, but not necessarily experience in method development or method transfer. This course guides attendees through each step of a process that leads to robust, efficient HPLC methods.
Understand the factors affecting the chromatographic resolution
Appreciate the advantages of small particle-size columns in terms of reduced peak width and better sample throughput
Have the knowledge required to design method conditions appropriate for hydrophilic and ionizable solutes
Understand common solute-stationary phase interactions mechanisms
Appreciate the effect of gradient delay volume on gradient separations
Know the important properties of common detector types
Be able to define the performance required of an analytical method before starting work on the development; Understand the influence of the sample matrix on method performance and robustness
Know how to approach the development of a gradient HPLC method in a logical, scientifically-sound manner
Appreciate the differences in chromatographic behavior between large and small molecules
Be able to evaluate methods prior to transfer and address method weaknesses in the transfer strategy
- Revision of chromatographic theory
Important chemical concepts (polarity, hydrogen bonding and acid/base theory)
Definitions of HPLC terms
The Van Deemter equation
Factors affecting resolution
Factors affecting resolution
Chiral separations
Column packing types and the effect of particle size
Strategies for ionisable compounds
Pump types and their effect on gradient separations
Effect of column temperature
Common detector types
- HPLC Method Development
Considerations before starting
Analyte properties
Matrix properties
Method performance requirements
Sample preparation
Options for different compound types
Retaining polar and ionisable compounds
Optimising resolution
Stationary and mobile phase chemistry
Stationary and mobile phase chemistry
Gradient separations
Detector selection
Derivatisation to improve detectability
Biochemical separations
Biochemical separations
- Transferring existing methods
Method transfer guidance – USP <1224>
Method changes allowed by compendial guidance (USP/EP)
C18 stationary phase equivalence – hydrophobicity and silanol activity/acidity
Method transfer strategies
Risk-based approach to method transfer acceptance criteria
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