An Introduction to the Principles and Practices of High Performance Liquid Chromatography

Presented by David Carr, Analytical Biotechnologies

A one-day short course introducing scientists to the basic concepts and operational principles of High Performance Liquid Chromatography with an emphasis on Reversed-Phase HPLC. The class makes extensive use of computer animation to explain basic principles, show instrument operation and to explain chromatography troubleshooting. Handouts printed in four colors aid in understanding the material presented.

Course Content

• Introduction to chromatography: A brief historical perspective leads into what chromatography is and how separations are performed.
• Chromatographic process: The chemical interactions behind chromatographic separations are explained in detail. This provides a basis for discussing the four modes of chromatography: reversed-phase, normal phase, ion exchange and size exclusion. The class focuses mainly on reversed-phase HPLC because of its dominant role in HPLC.
• Instrument operation: Using computer animation, instrument operation is explained and an example chromatogram is shown. Each part of an HPLC instrument is described in detail, including:

• The solvent delivery system which provides a constant flow of mobile phase.
• Sample injection.
• Connecting tubing and fittings.
• Commonly used detectors.

• Instrument operation: Using computer animation, instrument operation is explained and an example chromatogram is shown. Each part of an HPLC instrument is described in detail, including:
• Chromatograms: An example of a reversed-phase chromatogram is shown and significant aspects of the chromatogram such as column void volume and peak efficiency (theoretical plates) are explained.
• Columns: The heart of chromatography is the column that separates the sample constituents. Modern reversed-phase HPLC columns are explained and their make-up is described. Proper maintenance of columns is explained and cleaning procedures are described.
• Mobile phase: The mobile phase carries the sample components through the column and affects the retention and separation. The important attributes of the commonly used reversed-phase solvents are discussed and concepts that affect retention and separation such as solvent strength are explained. The class describes how to adjust solvent conditions to get satisfactory retention. Isocratic and gradient elution are explained.
• Instrument maintenance: Important steps to proper maintenance of an instrument are described
• Diagnosing and correcting chromatographic problems: Based on animated figures and the understanding of instrument operation previously discussed, problems that can occur during HPLC are explained and ways to avoid or correct these problems are discussed. Chromatographic problems such as tailing peaks are also discussed and ways to avoid problems are outlined. The three main indicators of chromatographic performance are described.

Course Outline

Morning

• An introduction to high performance liquid chromatography.
• Chromatographic interactions and the four modes of HPLC.
• Instrument operation including solvent delivery system, sample injection, fittings and tubing and detectors.

Afternoon

• Understanding and interpreting HPLC chromatograms
• Reversed-phase HPLC columns.
• Reversed-phase HPLC mobile phases
• Diagnosing and correcting instrumental and chromatographic problems.

About the instructor

David Carr has been involved in high-performance liquid chromatography since 1971. He worked with early instrumentation, helped design the first variable wavelength UV detector and has worked with all of the different types of columns used in HPLC over the past 35 years. David has been teaching the basic principles of HPLC for 35 years and has developed several classes for understanding the basic principles of HPLC and the important aspects of reversed-phase HPLC method development. He currently also teaches the class “In Search of Resolution”, a class focused on explaining in detail the important variables involved in reversedphase chromatographic separations, how each affects retention and resolution and how to most efficiently and effectively develop reversed-phase HPLC methods. David is very knowledgeable and experienced and is rated an excellent instructor by those who attend his classes.

Who Should Attend:

New chromatographers or current users who wish to review the basics.

Ordering Information:

Registration: In the USA and Canada, use the online registration form or contact training@dionex.com.

Schedule:
Dionex NACSC , Bannockburn, IL

August 19, 2008

November 11, 2008

Note: Attend “An Introduction to the Principles and Practices of HPLC” and “In Search of Resolution ” together, for the combined price of $950 per person.

Would you like to attend multiple courses? Make the most of your trip by attending more than one training class. Check out the complete Dionex NACSC training calendar for an overview of all courses offered.