Effective LC method development is crucial for achieving reliable, high-quality separations. This tutorial series will equip you with practical tools and insights to streamline the process, ensuring flexibility and precision in your workflows. From performance monitoring to optimizing mobile phases and column selection, you'll gain valuable knowledge to enhance your analytical capabilities.
The following modules are included in this series:
- Ensuring Proper System Performance Before Method Development: Discover a simple and easy performance monitoring process that is not tied to a specific assay, allowing more flexibility in testing.
- Selecting An Appropriate Method Development Approach: This module will look at the different method development approaches. The approach selected should be based on sample complexity and the need for a fast, traceable method development process.
- The Importance of Testing High and Low pH Mobile Phase Additives: Delve into the importance and limitations of using both low pH and high pH modified mobile phases as a part of method development. While switching between the two is highly beneficial from a selectivity standpoint, column stationary phases have limitations that analysts should be aware of.
- Differing Selectivity Obtained by Using Alternate Mobile Phase Strong Solvents: This module looks at the different selectivity that can be achieved by using alternate mobile phase strong solvents. By using methanol instead of acetonitrile, additional retention mechanisms can be realized promoting alternative selectivity, especially on certain stationary phases.
- Column Selection A Choice of Utmost Importance: Learn about the column stationary phases in more detail. We discuss particle morphology, particle type, and bonded ligands and provide recommendations for where each is most applicable.
- Instrument Contributions to Column Performance and Method Development: The final module covers the topic of instrument contributions to column and assay performance. Considerations of system dwell as it relates to column internal diameter are made.
Learning Outcomes
After completing this tutorial series, you will understand:
- The difference between a system suitability injection and a qualification check
- How to set up a system performance monitoring procedure
- The four main approaches to method development
- The benefits and limitations of using manual vs software-assisted method development
- How to use pKa to determine (approx.) at what pH a compound should be tested
- How alternating between low and high pH can drive selectivity in method development
- How using acetonitrile vs methanol will affect selectivity when using certain column chemistries, especially phenyl-hexyl and PFP phases
- How to select a column based on the analytes of interest instead of selecting columns based on past research
- How to build a panel of columns to assist with method development
- How the different systems affect method development
- How to properly select column configurations based on the system being used
Plus, download these additional resources
Wall Chart: Waters Column Selection Guide for Polar Compounds Wall Chart
Wall Chart : Waters Column Selection Guide for Polar Compounds Wall Chart
Wall Chart: Waters UPLC, UHPLC, and HPLC Column Selection and Mobile Phase Guide
Wall Chart: Simplified Column Selection and Method Development
Kenneth Berthelette
Senior Scientist, Waters Corporation Chemistry and Consumables Group
Ken earned his Bachelor's of Science degree in Professional Chemistry from Bridgewater State University, Bridgewater Massachusetts, in 2010. He has been with Waters Corporation since March 2012 and has worked on several product launches including CORTECS columns, UPC2 Torus and Trefoil columns, and the Atlantis Premier BEH Z-HILIC column. He changed job functions in 2021 and now supports Chemistry Aftermarket initiatives. His focus on applications and customer education has led to several seminars and webinars on topics including method development, method modernization and transfer, HILIC separation techniques, and LC system troubleshooting.
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