In Search of Resolution - A Practical Approach to Reversed-phase HPLC Method Development

Presented by David Carr, Analytical Biotechnologies

A one-day short course focused on revealing the important factors involved in reversed-phase chromatographic separations, how each affects retention and resolution and how to most efficiently and effectively develop reversed-phase HPLC methods.

Reversed-phase method development is looked upon as a mystery by many chromatographers. Resolution seems to be a hidden treasure which either requires too much time to find or entirely eludes the earnest seeker. The class is presented as a search for the treasure of resolution. A map is clearly

laid out guiding the chromatographer through each step of the pathway that leads to resolution. Those attending the class will learn the important principles which affect resolution and will be presented with a systematic, efficient approach to developing reversed-phase HPLC methods.

Check out our April “3 for 2” promotion.

Course Content

• Introduction to Resolution. The concepts of efficiency, retention factor and selectivity are discussed in detail as they relate to chromatographic resolution. The way to optimize each parameter and its role in reversed-phase resolution is explained.
• Reversed-Phase HPLC Columns. The characteristics of reversed-phase columns are explained as they contribute to retention and resolution.
• The matrix and the nature of the stationary phase
• How hydrophobic, polar and ionic interactions contribute to and affect reversed-phase
• retention and selectivity.
• The type of stationary phases available and how each affects selectivity.
• How to evaluate and compare columns in order to select the best column for a given separation.
• Reversed-Phase HPLC Mobile Phases. The characteristics of mobile phases used in reversed-phase HPLC are explained.
• How to find the best solvent concentration for a method
• How solvent type and concentration affects selectivity and why
• How to best perform gradient elution
• How gradient elution affects selectivity.
• How the pH of the mobile phase is used to optimize the separation of basic and acidic compounds.
• Temperature. The affect of column temperature on retention and selectivity is described.
• Method development. The principles of reversed-phase method development are brought together in a systematic approach to developing effective, robust reversed-phase HPLC methods in the most efficient manner.

Course Outline

Morning

• An introduction to the principles of chromatographic resolution.
• Reversed-phase HPLC columns

Afternoon

• The mobile phase: Solvents used and their characteristics.
• How the solvent affects retention and selectivity. The affect of solvent strength and type.
• Gradient elution and how it affects retention and selectivity
• The use of pH to control the retention of acids and bases.
• The affect of temperature on reversed-phase separations
• A systematic approach to developing reversed-phase HPLC methods

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 “An Introduction to the Principles and Practices of HPLC”, a class which introduces the basic principles of HPLC and explains instrument operation, maintenance and troubleshooting.. David is very knowledgeable and experienced and is rated an excellent instructor by those who attend his classes.

Who Should Attend:

Chromatographers wishing to improve their knowledge of reversed-phase method development.

Ordering Information:

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

Schedule:
Dionex NACSC, Bannockburn, IL

August 20, 2008

November 12, 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.