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Lecturer(s)
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Ševčík Juraj, prof. RNDr. Ph.D.
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Petr Jan, doc. RNDr. Ph.D.
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Course content
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Migration of ions in electrical field. Ion mobility. Limit, actual and effective mobility of ions. Types of electromigration methods. Electrophoresis and electroosmotic flow. Electrokinetic zeta-potential, electric double-layer. Electrophoretic velocity and its influencing, mobilities and electroomosis. Zonal electrophoresis. Band broadening and suppression of diffusion by stabilization of zone. Analytical and preparative slab gel electrophoresis. Capillary zone electrophoresis. Efficiency and resolution. The influence of Joule heating, electroosmosis, adsorption, electromigration dispersion on efficiency. The influence of chemical modifications of capillary, radial voltage. The choice of running electrolyte, voltage. Instrumentation. Separation of neutral compounds. Micellar electrokinetic chromatography. Special influencing of effective mobilities. Inclusion complexation, crown complexes, afinity complexes, enantiomeric separation. The dynamic changes of composition of running electrolyte during separation processes. Izoelectric focusation, characteristics of focusation. Fucusation in gradient of pH and in gradient of ligand. Self-sharpen effect, the condition of isotachophoresis. Kohlrausch regulation function - the condition of continuity. Leading and terminating electrolytes. The prediction and mathematic optimization of separation conditions. The connection of ITP and CZE. On-line preconcentration techniques. Application of methods on low-molecular compounds and macromolecules.
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Learning activities and teaching methods
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Lecture
- Attendace
- 13 hours per semester
- Homework for Teaching
- 13 hours per semester
- Preparation for the Course Credit
- 34 hours per semester
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Learning outcomes
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The aim is to become familiar with the principles and instrumentation in electromigration methods.
Describe the basic principles of ionic species migration in the electric field. Define the relations between the composition of background electrolyte and migration of ionic species. Describe the basic instrumentation for the electrophoretic separation based on the chip technology, capillary and slab gel technology.
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Prerequisites
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The knowledge of basics of analytical chemistry and instrumental methods of analytical chemistry is recommended.
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Assessment methods and criteria
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Oral exam
student has to pass successfuly an oral colloquium- discussion on optional two topics
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Recommended literature
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1. J. P. Landers, Handbook of capillary electrophoresis, CRC Press (1997)..
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2. D. R. Baker, Capillary electrophoresis, Wiley and sons, (1995)..
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3. F. Fore, L. Křivánková, P. Boček, Capillary zone electrophoresis, Wiley and sons, (1993)..
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4. R. Westermeier, Electrophoresis in Practice, A Guide to Methods and Applications of DNA and Protein Separartions, Wiley and sons, (2005)..
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5. R. Westermeier, Electrophoresis in Practice, A Guide to Methods and Applications of DNA and Protein Separartions, Wiley and sons, (2005)..
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