Mass Spectrometry and Molecular Analysis

Chromatographic and electrophoretic separation techniques I

Integrated course, 4.00 ECTS


Course content

Foundations of chromatographic techniques:
- Taxonomy of chromatographic procedures
- principles of chromatographic separation
- liquid chromatography
- instrumentation
- important parameters
- Van Deemter equation
- stationary phases
- mobile phases
- reversed-phase chromatography
- normal-Phase chromatography
- ion exchange chromatography
- hydrophilic interaction chromatography
- chiral chromatography
- affinity chromatography
- gel-permeation chromatography
- principles of isocratic separation and gradient separation
- detectors
- Gas chromatography
- instrumentation
- important parameters
- Raoult's law
- separation columns
- carrier gases
- retention mechanisms
- detectors
- principles of temperature dependent separation of volatile and semi-volatile compounds
Foundations of electrophoretical techniques:
- principles of electrophoretical separation
- capillary zone electrophoresis
- instrumentation
- retention mechanisms
- detectors

Selection and application of chromatographic and electrophoretic separation techniques in chemical analysis, theory and practice of opitimization of separation parameters

Learning outcomes

The students understand the fundamental mechanisms and formal relationships between chromatographic, electrophoretic and spectrometric techniques. In the field of chromatographic methods they can characterize chromatographic processes with regard to the classification of liquid, gas, gas/liquid, thin-layer chromatography (LC, GC, GLC and DC) and capillary electrochromatography (CE). They know the basic technical structure of LC and GC systems and can describe the individual function modules. According to the separation criteria they are able to differentiate between normal phase (NP - LC), reversed phase (RP - LC), ion exchange (IEX- LC), size-exclusion chromatography (SEC - LC) and hydrophilic interaction chromatography (HILIC) and can apply them goal-oriented. They can successfully apply criteria for the choice of carrier materials and the associated separation method. In GC and GLC the students can explain the fundamental principle of separation of analytes and can choose an appropriate separation column and the associated parameter regime. In electrophoretic procedures, the students can explain the physical principles with regard to the separation criteria of molecular weight and the associated hydrodynamic factor and the charging of analytes and can choose a suitable technology in a specific situation. They know about native and denaturing methods and are able to define suitable separation conditions with regard to the choice of the required puffer systems and the associated pH value ranges. In the field of spectrometric procedures and techniques the students develop competence in the theory of UV/VIS IR-, Raman-, fluorescence- and mass spectrometry. They can differentiate between electron excitation spectra and molecular vibration spectra. They are familiar with the principle of fragmentation of molecules in mass spectrometry. They are able to to choose the suitable procedure of a given task and to plan and carry out the analytical processes. They know how to interpret appropriate spectra, and, thus, can evaluate and interpret the results of their analyses.