Syllabus
1. Introduction into separation methods, basic terms and theory of separation processes, classification of separation methods, interaction forces in separations processes.
2. Extraction, classification of extraction methods, extraction of inorganic, organic substances and ionic chelates, reextraction.
3. Solid phase extraction (SPE) and solid phase microextraction (SPME), principles, instrumentation and utilisation.
4. Supercritical fluid chromatography (SFE), supercritical medium, choice of supercritical fluid and modificator, types of analyte collection.
5. Affinity chromatography (AC), principles, affinity of ligands to macromolecules, binding of affinants to solid substrates, solid substrates and their selection, elution techniques.
6. Ion exchange chromatography (IEC), ionex types and their characterisation, application to inorganic and organic substances.
7. Gel permeation chromatography (GPC), molecular size exclusion effect, types of gels, separation of group of substances and desalination, fractionation and determination of relative molecular weight.
8. Gas chromatography (GC), adsorption and absorption, sorption isotherms, dissolution, rate and plate theories of GC, efficiency of GC, detectors, packed and capillary columns, injection techniques, stationary phases, qualitative and quantitative analysis, calibration procedures.
9. Chromatograph for high performance liquid chromatography (HPLC). Stationary and mobile phases, affinity, distribution and retardation. Thermodynamics and kinetics of separation. Retention quantities. General equation of chromatography. Zone broadening causes. Separation efficiency and resolution. Analytical information in chromatogram. Pumps, injection valves, columns and detectors. Stationary and mobile phases in LSC, LLC, IEC and GPC. Applications in HPLC.
10. Paper chromatography (PC). Thin layer chromatography (TLC). Development techniques. Detection and identification. Retardation factor. Densitometer. HPTLC. Applications in PC a TLC.
11. Electromigration methods. Electrophoretic migration and mobility. Electroosmotic flow and mobility. Electrical double layer and zeta-potencial. Velocity and temperature profile. Equipment for capillary zone electrophoresis (CZE). Separation mechanism in CZE. Electropherogram. Calculation of mobilities. Weak electrolytes. Sample injection. Detection. Analytical information in electropherogram. Applications in CZE.
Annotation
The course of separation methods gives compendium on modern separation methods from the theoretical and practical point of view. The course explains the general theory of separation and a summary of physico-chemical principles such as transport phenomena, solubility, adsorption, ion exchange, exclusion effect, biological recognition. Theory of chromatography and of electromigration. Systematic description of important separation techniques. The course ends through a written exam.
Learning outcomes: After completing the course, the student will be able to define basic terms, clarify the theory of separation processes, differentiate interaction forces in separation methods, perform extraction, explain solid phase extraction, clarify supercritical fluid extraction, define affinity chromatography, define ion exchange chromatography, define gel permeation chromatography, perform gas chromatography, explain supercritical fluid chromatography, perform liquid chromatography, perform paper chromatography, perform thin-layer chromatography, explain electromigration, explain the overview and division of electromigration methods and perform capillary electrophoresis.