A comprehensive course in the systematic development of liquid chromatography separations using QbD principles. Normal phase is less used today than reversed-phase, but it can be a powerful tool to separate mixtures containing structural isomers and other compounds of similar polarity that are difficult to separate by reversed-phase techniques. We discuss the normal-phase retention mechanism, both for bare silica and bonded phase columns.
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.
This course covers:
Normal phase is less used today than reversed-phase, but it can be a powerful tool to separate mixtures containing structural isomers and other compounds of similar polarity that are difficult to separate by reversed-phase techniques. We discuss the normal-phase retention mechanism, both for bare silica and bonded phase columns. Alternatives to the normal phase are considered. HILIC (hydrophilic interaction chromatography) may be the fastest growing mode of chromatography today. HILIC uses a normal-phase retention mechanism in a mobile phase that might be considered reversed-phase, such as acetonitrile-buffer mixtures. We look at the retention mechanism involved with HILIC and how to take advantage of it to develop HPLC and UHPLC methods in an efficient manner.
- Normal Phase
• Problems & Alternatives
• What Is It?
• How Does It Work
• Method Development
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 true about normal phase separations?
- What best describes the normal phase retention mechanism?
- What is normal phase retention?
- What is true about solvent strength in normal phase?
- What is the role of water in normal-phase separations on bare silica?
- Why would someone choose a polar bonded phase instead of bare silica for normal phase?
- Why are polar modifiers, such as ACN or MeOH, used in normal phase on bare silica?
- What is HILIC?
- When defining a process as normal phase (NP) vs. reversed phase (RP), how does the mobile phase polarity differ?
- What is the role of water in HILIC?
- How do we explain HILIC retention?
- Why is ACN the preferred organic solvent for HILIC?
- What is the role of mobile phase pH in HILIC retention?
- Which mechanism is most predominant in determining HILIC retention?
- What is the best strategy for choosing a HILIC column?
- Which is a good strategy for developing a HILIC separation?
- Which strategy is appropriate for HILIC 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.