High Performance/Pressure Liquid Chromatography (HPLC) in Theory and Practice

Content & Learning Objective

Chromatographic separation has become the most powerful tool in analytical laboratories. High Performance/Pressure Liquid Chromatography (HPLC) and the more recent development of Ultra-high Performance/Pressure Liquid Chromatography (UHPLC/UPLC) are essential tools for the analysis of anything from a single compound up to complex mixtures. In this course you will learn the basics of chromatographic separations, how each part of the HPLC system functions, and the development of chromatographic methods and their applications.

In the theory section we will cover:

  • Introduction: Applications of HPLC in the laboratory
  • Basics of chromatographic separations
  • The HPLC system and the function of its components
  • Development of a RP method
  • Development of other separation methods
  • Data analysis and report generation
  • Transfer of HPLC Methods to UHPLC/UPLC

In the practical section we will cover:

  • Step-by-step through the HPLC system (function of the components, trouble finding and troubleshooting
  • Analysis of a single compound by RP chromatography
  • Separation of substance mixture
  • Optimization of the separation, generation of a gradient
  • Problems with complex mixtures
  • Data analysis and documentation

Target Group

Technical and scientific staff with good basic knowledge of cell culture and flow cytometry. Basic familiarity with flow cytometer is advantageous but not required for attending this course.


Dr Manfred Raida studied chemistry at the FU Berlin and completed his PhD at JW Goethe University and the MPI for Biophysics, Frankfurt, Main. After his PhD he worked as a research fellow at DIAGEN (today QIAGEN, Hilden); as group leader for peptide analysis at the Lower Saxony Institute for Peptide Research (IPF, Hannover); European Support Manager for Mass Spectrometry at Amersham (today GE, Freiburg) and as senior research fellow at Cellzome (today GSK, Heidelberg). Following this, he worked as group leader in Mass Spectrometry at the Experimental Therapeutics Centre (A*STAR, Singapore) and as senior research fellow at the National University of Singapore, Singapore Lipidomics Incubator. His fields of expertise are separation science, mass spectrometry, proteomics, lipidomics, metabolomics and analysis of small molecules in drug development processes.