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.

Pricing: Starting USD 2995
Duration: >1 day
Coming Soon

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

Youtube ID: TQbWtrcim3A

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.

Course Outline

  • Topics Covered

    Introduction and Definitions



    HPLC Theory


    HPLC Columns

    Reversed Phase of Ionic Samples

    Quantitative Analysis

    Normal Phase

    Hydrophilic Interaction (HILIC)

    Ion Exchange Chromatography

    Size Exclusion Chromatography

    Method Development

    Preventative Maintenance

    Troubleshooting Exercises

Dates, Locations, and Prices

Course fee includes tuition, lecture and laboratory materials, morning and afternoon snacks, and all lunches.

View complete details about lodging, local dining options, and available transportation here.

The course runs from 9:00 AM to 5:30 PM each day, class ends at 3:00 on Friday

Check in opens at 9:00 AM on the first day of the course.


Axion Analytical Labs & Training Institute
14 N Peoria St #100
Chicago, IL 60607

Early Bird Full Price
Member(Premium Package) $2995
Member or Associate(Standard or Basic Package) $3195
Non-Member $3195