Conducted by John W. Dolan, Ph.D.
Author of LCGC's popular LC Troubleshooting Column

As the most widely used analytical technique in the world, high performance liquid chromatography (HPLC) is a tool touching the lives of many laboratory workers. Reversed-phase HPLC accounts for 70-80% of all applications, so understanding its fundamentals is important if you want to take full advantage of this powerful technique. This Audio Seminar CD covers the basics of the reversed-phase separation, how to control retention and selectivity, and the role the column plays in the process. Whether you are an experienced user that needs a tune-up or a newcomer to HPLC, this teleseminar will provide a solid foundation for understanding what is happening behind the scenes in the HPLC process.

Over the last 30 years, Dr. Dolan has designed LC equipment, developed methods, managed a contract research laboratory, taught thousands of students in LC techniques, and written over 100 papers on LC. His popular LC Troubleshooting column in LC/GC has been the go-to article each month for more than 20 years. He draws on this experience and his expertise in HPLC for this Audio Seminar CD.

Specific areas that will be covered include:

• How does reversed-phase work?
• How can I use solvent strength to control the separation?
• How do temperature, pH, solvent type, and ion pairing influence the chromatogram?
• What difference will the choice in column packing make?
• Which variable should I choose first?

This Audio Seminar CD is presented by Dr. John W. Dolan, worldwide expert on method development and troubleshooting liquid chromatography. In this unique teleseminar format, lasting just one hour, Dr. Dolan will cover the key aspects of reversed-phase chromatography. You will learn how each variable in the process works and how to choose which one to try first for best results. The understanding you will gain will help you make better use of your time when working with reversed-phase HPLC methods.

Reversed-Phase Separation in HPLC

Part I Reversed-Phase and solvent strength

• A review of basic measeurements
• Solvent strength and retention
• Solvent strength and selectivity

Part II pH, solvent type, and other variables

• Temperature, retention, and peak spacing
• The effect of solvent type and solvent blends on selectivity
• pH - a powerful tool, but very touchy
• Ion pair - a last resort?

Part III The role of column type in selectivity

• Bonded phase preparation
• Bonded phase selectivity
• Which variable to use first?

John W. Dolan, Ph.D.

Dr. John Dolan is most widely known as the author of LCGC’s popular LC Troubleshooting column. As editor of this column for over 20 years, Dr. Dolan has contributed more than 200 installments of practical advice to practicing chromatographers. John has written over 100 technical papers, many of which support the development and practical application of gradient elution LC. His book (with Lloyd Snyder), Troubleshooting LC Systems, is a standard reference in laboratories worldwide. He is the 2002 recipient of the prestigious Palmer Award given by the Minnesota Chromatography Forum.

Dr. Dolan managed a regional contract research laboratory for eleven years for LC Resources and later Bioanalytical Systems, Inc. The lab, which he helped found in 1988 specializes in the development, validation, and application of LC-MS/MS methods to determine pharmaceutical compounds in biological matrices. In addition, he is one of the founders of LC Resources, Inc., a company dedicated to training chromatographers around the world. As an instructor for LCR and the American Chemical Society, John has instructed more than 10,000 students in practical LC techniques.