In-Person High Performance Liquid Chromatography: Fundamentals, Troubleshooting, and Method Development
Gain practical laboratory experience in HPLC and learn the fundamentals needed to understand the techniques and instrumentation involved in this powerful analytical tool.
About the Instructor(s)
Lee N. Polite
President, Axion Analytical Labs, Inc
Dr. Lee N. Polite is the president and founder of Axion Analytical Labs, Inc. He has taught chromatography to customers from every major pharmaceutical, chemical, and petroleum company in the U.S.
About the Course
This course is designed for beginning- and intermediate-level users in HPLC who want practical laboratory experience. Lectures are supplemented by problem sets, slides, and video presentations.
Key topics include: basic instrumentation for HPLC; detectors, including UV/VIS, photo diode array, mass spectrometer, and fluorescence; column selection and optimization; qualitative and quantitative analysis; and troubleshooting HPLC systems
- HPLC Overview–introduction to HPLC; instrumentation overview; detector details
Laboratory: basic HPLC hardware familiarization
- Separation Fundamentals and Mobile Phase–practical HPLC theory; important HPLC parameters; the mobile phase
Laboratory: mobile phase strength, flow rate, and temperature
- Separation Modes and Columns–reversed phase; normal phase; ion exchange; size exchange
Laboratories: pH effects in reversed phase and column dimensions
- Quantitative Analysis and Troubleshooting–qualitative and quantitative analysis; LC/MS; hardware troubleshooting; separation troubleshooting
Laboratory: internal vs. external standard; hardware troubleshooting.
- Gradients and Method Development–gradients; fundamentals of method development
Laboratory: develop a method to separate a completely unknown mixture.
What You Will Learn
Each attendee will leave the course with the ability to:
- Understand how each HPLC parameter affects separation
- Build a calibration table to quantify samples
- Develop and optimize HPLC methods from scratch
- Cut analysis times in half
- Speak as an authority in chromatography
- Troubleshoot and fix HPLC issues
Who Should Attend
Beginners and intermediate-level HPLC users who want practical laboratory experience.
No prior experience in advanced mathematics, theoretical chemistry, or physics is required for this course.
Some experience in HPLC or introductory course work in organic chemistry is desirable but not necessary.
Introduction and Definitions
Reversed Phase of Ionic Samples
Hydrophilic Interaction (HILIC)
Ion Exchange Chromatography
Size Exclusion Chromatography