Course: Separation Methods

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Course title Separation Methods
Course code TUFMI/TE9SM
Organizational form of instruction Lecture + Tutorial
Level of course Master
Year of study not specified
Semester Winter
Number of ECTS credits 4
Language of instruction English
Status of course unspecified
Form of instruction Face-to-face
Work placements This is not an internship
Recommended optional programme components None
Course availability The course is available to visiting students
Lecturer(s)
  • Ingr Marek, RNDr. Ph.D.
Course content
1. Introduction to separation methods, principles of separation. 2. Intermolecular interactions and properties of compounds stemming from them. 3. Processing of complex biological material, basic separation methods (filtration, extraction, precipitation, salting). 4. Electromigration separation methods - principle of zone electrophoresis and isotachophoresis. 5. Survey of capillary and gel electrophoresis methods and their use. DNA sequencing. 6. Sedimentation methods as a tool of analysis of natural compounds - sedimentation velocity in a gravity field and ultracentrifuge. 7. Sedimentation equilibrium, dissociation equilibrium of proteins, isopycnic centrifugation. 8. Chromatographic methods - basic chromatographic principles, analysis of results. 9. Isocratic chromatography methods, gel permeation chromatography - principles and examples of use. 10. Gradient chromatographic methods - ion exchange chromatography, hydrophobic chromatography, reverse phase chromatography, affinity chromatography - principles and examples of use. 11. Principles of detection in chromatographic and electromigration methods - basics of spectrophotometry, fluorometry, refractometry. 12. Detection methods in gas chromatography, mass spectrometry. 13. Methods of natural-compounds analysis - concentration assays, amino acid analysis, protein sequencing. 14. Blotting, PCR and qPCR, immunological methods, flow cytometry.

Learning activities and teaching methods
Lecturing, Simple experiments
Learning outcomes
The aim of the course is to give the students the information about the methods of isolation of biologically active compounds, especially proteins, from biological materials. It consists of both theoretical and practical training of the chromatographic, electrophoretic, sedimentation and other techniques. The stress is put especially on understanding the principles and applications of the methods in complex purification projects and their use for analytical purposes. The students will also learn the analytical applications of the discussed techniques.
Students understand physical-chemical principles of methods of isolation of proteins, DNA and other natural and non-natural compounds. They are able to design an experimental method to isolate raw material to obtain pure substance and they are able to characterize and quantify the final product.
Prerequisites
Knowledge of physical and analytical chemistry at the level of bachelor study program, basics of organic and inorganic chemistry, biochemistry, physics and mathematics. Knowledge of the basic principles of work in laboratories.

Assessment methods and criteria
Grade (Using a grade system), Oral examination, Essay

Finishing the subject: Credit: performing the laboratory exercises and writing a seminar work on the topic of one of them. Examination: knowledge of the discussed curriculum content, oral examination. The result of a subject examination is expressed on a six-point scale: A "výborně" (i.e. "excellent"), B "velmi dobře" (i.e. "very good"), C "dobře" (i.e. "good"), D "uspokojivě" (i.e. "satisfactory"), E "dostatečně" (i.e. "sufficient"), F "nedostatečně" (i.e. "fail").
Recommended literature
  • I.D.Wilson. Encyclopedia of Separation Science. New York, 2000. ISBN 9780122267703.


Study plans that include the course
Faculty Study plan (Version) Category of Branch/Specialization Recommended year of study Recommended semester