A comprehensive course in the systematic development of liquid chromatography separations using QbD principles. How can we take advantage of pH to improve the separation and what are the pitfalls when we do use pH for this purpose? Why is temperature more important when we have ionic compounds present? 

This course is designed for practical workers in the laboratory who have the responsibility for designing new HPLC or UHPLC methods or transferring methods between laboratories. Experienced workers will get the most out of this class, but because it builds from the fundamentals, anyone with experience in HPLC or UHPLC will gain valuable knowledge.

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This course covers:

How can we take advantage of pH to improve the separation and what are the pitfalls when we do use pH for this purpose? Why is temperature more important when we have ionic compounds present? How can ion pairing be used to separate some samples that cannot be separated by pH alone? What is mixed-mode chromatography and how can it be used?

  • The Role of pH 
  • Tailing Peaks 
  • Ion Pairing – Controlling Retention
  • Ion Pairing – Special Considerations 

By attending this online training course you get full access to the 38 video modules and approximately 12.5 hours of instruction. You also get handouts containing copies of all of the approximately 520 PowerPoint slides used in the class. These are arranged for easy note-taking while you view each module and give you a valuable resource for future reference.

This module has a complementary quiz. By correctly answering the quiz questions for all of the modules as part of the course you will be able to download your certificate of completion. Types of questions to expect are:

  • What is the effect of the mobile phase pH in reversed-phase HPLC?
  • Why would pH adjustment be a good strategy to obtain a separation of a mixture of neutral and ionizable compounds?
  • How to choose a buffer?
  • What is peak tailing in reversed-phase HPLC?
  • What affect the peak spacing when ionic compounds are present?
  • What is the purpose of adding triethylamine (TEA)?
  • What is true about peak tailing?
  • Why can temperature changes give similar results to changes in mobile phase pH?
  • What is ion pairing?
  • What would you expect to happen as you increase the concentration of negatively charged ion-pair reagent in the mobile phase?
  • When the concentration of a negatively charged ion-pair reagent is kept constant and the percent organic in the mobile phase is increased, what do you expect to happen to retention?
  • How does column temperature affect ion pairing separations?
  • What generalizations can be made about mixed-mode chromatography?

 

Analytical Training Solutions, brought to you by Separation Science, is the leading global portal for fundamentals, best practice, troubleshooting and method development training for chromatographic and mass spectrometric techniques. Comprehensive, self-paced online courses and validated learning provides a unique education resource for analytical chemists. Currently, we offer HPLC training, LC-MS training, GC training and GC-MS training.

 

Published  Mar 27, 2023

Home 9 Techniques 9 Liquid Chromatography 9 Reversed-Phase HPLC of Ionizable Analytes