HPLC for the Pharmaceutical Chemist

(One-day short course)

Course summary:

This course is taught at the intermediate level and is intended to serve chemists working in the pharmaceuticals and biotechnology industries, though the topics covered are of equal benefit to chemists working in other fields.  The audience should consist of persons who routinely work with HPLC and/or other chromatographic techniques, and whose understanding of chromatography is at the intermediate level.

Topics include all stages of method development: sample handling, selection of the separation technique, and column and mobile phase selection. Special attention will be given to the most popular modes of liquid chromatographic separation, LC-MS, CE and CEC as well as on specific techniques such as separation of peptides and proteins, and of ionic and ionizable components (pharmaceuticals). Participants are also encouraged to bring their own separations problems for discussion.

Who will benefit from the course:

The course is appropriate for individuals with prior HPLC experience, though beginners are most welcome.  The course was developed specifically for those working in the pharmaceuticals industry research chemists, laboratory technicians, biochemists, and biologists searching for practical solutions to separations problems.

Course agenda:

8:30        Introduction

The broader approach to developing HPLC analysis strategies

9:00        Sample handling and pretreatment

                        Principles of sample preparation in view of the ensuing analytical technique.
                        Matrix effects; extraction modes; diluent selection

9:50         Coffee

10:00     Reverse phase HPLC column selection and pharmaceutical method development

Main column parameters, system suitability, surface chemistry,  eluent type and composition effects

11:00     HPLC techniques for the analysis of ionized or ionizable analytes

General approach to selection of a chromatographic technique.  Principles and applications of  ion suppression HPLC, ion-exchange HPLC, ion chromatography and ion pair HPLC.  Specific modes of detection for carbohydrates, etc.

12:00     Lunch

13:00     Chromatographic separations of proteins and peptides

Special considerations in protein and peptide separations.  Selection of an appropriate HPLC mode, optimization of analysis parameters, and recovery of analyte.  Specific modes of detection for proteins and peptides.

14:00     The effect of eluent pH on reverse phase chromatographic behavior of pharmaceutical compounds

Fine tuning of the HPLC separation with pH adjustment. Buffer concentration effect. Influence of counteranions on the retention of basic pharmaceutical components

15:00       Break

15:15     Principles of LC-MS analysis

Major types of LC-MS interfaces, operation principles. Strategy of HPLC method development suitable for LC-MS, sensitivity enhancement.

16:15     Emerging techniques in liquid phase separations CE and CEC.