A comprehensive course in the systematic development of liquid chromatography separations using QbD principles. We concentrate on reversed-phase separations in this course, because the majority of HPLC and UHPLC methods use this technique. How does reversed-phase work? 

This course is designed for practical workers in the laboratory who have 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.


This course covers:

We concentrate on reversed-phase separations in this course, because the majority of HPLC and UHPLC methods use this technique. How does reversed-phase work? A homework exercise is given to help reinforce some of the relationships between solvent strength, retention, selectivity, and resolution. We show how you can choose a few key experiments and leverage the data into an understanding of how the separation will appear under a wide range of conditions. Why are different solvents used, and how do they differ in their properties and influence the separation? What is the role of temperature on the separation?

  • Solvent Strength & Retention 
  • Discussion of Workshop Results 
  • Temperature & Solvent-Type Selectivity 

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:

  • Which generality is true about reversed-phase HPLC?
  • Why is a 10,000 plate column a good starting point for most reversed-phase method development projects?
  • What does “Rule of Three” say?
  • What happens to the changes in the mobile phase percent organic (%B) in reversed-phase HPLC?
  • Which is true about reversed-phase retention and resolution?
  • What is true about the influence of the column on an existing isocratic separation?
  • What is the role of column temperature in reversed-phase HPLC?
  • What is the effect of changing solvents in reversed-phase HPLC?
  • Why is it a good idea to explore solvent strength changes before solvent type changes?
  • Why is ACN usually chosen over MeOH as the starting solvent for reversed-phase method development?


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.


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