Introduction to Practical HPLC Course - 2 days

Course duration:

Course content:

• History of HPLC
• Basic theory of HPLC
• Brief introduction to manual injection valves, isocratic pumps and UV detection
• Make injection and run basic HPLC chromatogram
• Effects of changing eluent composition, flow rate, column and detector
• Making up an accurate mobile phase composition and degassing
• Make flow path connections
• Basic precautions to avoid problems
• HPLC solvent selection
• Column efficiency and capacity
• Normal phase versus reversed phase analysis
• Introduction to chromatographic interpretation
• Introduction to sample preparation
• Introduction to autosamplers
• Safety: Working with small particle size materials, high pressures, UV radiation and electronic safety.

Working in pairs, we take an analytical method for the analysis of five sample components and set up a data system from scratch, working out the peak identification, the retention times and appropriate time windows for each peak. We decide on integration parameters and create a Peak Table. We then weigh out standards and make up dilutions for a five point calibration, and run the standards in duplicate to calibrate the method. Once we have checked the calibration we set the most appropriate calibration curve, decide whether to include the origin and then run unknown samples using the method we have created.

Assessment

Presentation of Course Certificates

By the end of the course you should know enough to:

• Make up a sample.
• Make up a mobile phase.
• Switch on and operate an isocratic HPLC system.
• Make a manual injection.
• Understand the use of autosamplers.
• Follow a basic HPLC method.
• Operate an HPLC system safely.
• Replace a column.
• Know if the chromatogram is integrated correctly.

Course Dates in 2010