PřF:C8022 Separation Methods B - Course Information
C8022 Separation Methods B
Faculty of ScienceSpring 2008
- Extent and Intensity
- 2/0/0. 2 credit(s) (plus extra credits for completion). Recommended Type of Completion: zk (examination). Other types of completion: k (colloquium).
- Teacher(s)
- doc. Mgr. Jan Havliš, Dr. (lecturer)
- Guaranteed by
- doc. Mgr. Jan Havliš, Dr.
Department of Experimental Biology – Biology Section – Faculty of Science - Timetable
- Tue 8:00–9:50 AK1
- Prerequisites
- C7021 Separation Methods A
C3100 + C4050 Analytical chemistry (at least C1660) C7021 Separation methods A - Course Enrolment Limitations
- The course is also offered to the students of the fields other than those the course is directly associated with.
- fields of study / plans the course is directly associated with
- there are 9 fields of study the course is directly associated with, display
- Course objectives
- Separation of macromoleculs (gel permeation chromatography (GPC), light scattering, hydrodynamic chromatography (HDC), field-flow frakcionation (FFF)) + Gas chromatography (GC), theory and praxis. + Electromigration methods = capillary zone electrophoresis (CZE), izotachophoresis (ITP), micelar electrokinetic chromatography (MEKC), gel capillary electrophoresis (GCE)
- Syllabus
- Separation of macromolecules
- Macromolecules: synthetic and biological. Radius of gyration. Relative molecular mass, polydispersity. Optical detection, viscosimetry. Ultracentrifugation, gel capillary electrophoresis. Matrix assisted laser desorption ionization-time of flight (MALDI-TOF). Separation methods for macromolecules: Gel permeation chromatography (GPC), Hydrodynamic chromatography (HDC), Field-flow fractionation (FFF). Special detection techniques. Static ligth scattering (LS), Low-angle laser ligth scattering (LALLS), multi-angle laser light scattering (MALLS). Hyphenation of separation and detection techniques.
- Gas chromatography
- Principles of gas chromatography, diffusion in gas phase, description of compound distribution between phases and character of compounds interaction in stationary and mobile phases. The influence of temperature and pressure of gas characteristics in gas chromatography. Basic components of gas chromatograph, sample injection. Thermo conductivity detection in gas chromatography (katharometer). Flame ionization detection (FID). Electron capture detector (ECD). Photoionisation detector, conductivity detector. Types of columns in gas chromatography, stationary phase support. Capillary columns in gas chromatography, influence of inner diameter of capillary and the film thickness on separation. Adsorption gas chromatography - common adsorbents. Adsorbed and covalently bonded stationary phases in gas chromatography - their properties, preparation and selection. Requirements for gas chromatography columns. Gas chromatography with programmed temperature - purpose, temperature influence on separation and characterization of retention.
- Electromigration methods
- Types of molecules that could be analyzed by capillary electrophoresis. Proteins. Polypeptides charge. Electrophoretic titration curve. Physical principles of capillary electrophoresis - electrophoretical mobility. Electroosmotic flow - electroosmotic pump, electroosmotic flow modificators - surfactants. Techniques of capillary electrophoresis: capillary zone electrohoresis (free solution capillary electrophoresis), capillary izotachophoresis (CITP), Capillary Gel Electrophoresis, Micellar electrokinetic chromatography, capillary electrochromatography, capillary izoelectric focusing, discontinual electrophoresis. Basics of instrumentation - experimental parametrs: capillary, temeprature control, detectors (indirect detection), sample introduction (injection), reduction of band broadening. Optimization: hea dissipation, electroosmotic flow, oncaillary detection, molecular diffusion, band broadening due to injection. Evaluation of electrophoretic efficiency, advantages od capillary electrophoresis. Evaluation of electropherograms: qualitative and quantitative. Limit of detection. Validation of methods of CE. SDS - Page electrophoresis, capillary ion electrophoresis (CIE), indirect detection, chiral separation, capillary electrohoresis in nonaqueous solvents, DNA analysis, laser induced fluorescence in capillary electrophoresis, capillary electrophoresis - mass spectrometry
- Literature
- CHURÁČEK, Jaroslav and Pavel JANDERA. Separace látek : kapalinová vysokoúčinná kolonová chromatografie. 1. vyd. Praha: SNTL - Nakladatelství technické literatury, 1981, 140 s. info
- CHURÁČEK, Jaroslav. Analytická separace látek. 1. vyd. Praha: Státní nakladatelství technické literatury, 1990, 384 s. ISBN 80-03-00569-8. info
- POPL, Milan and Jaroslav KUBÁT. Separace látek. 3. přeprac. vyd. Praha: Vysoká škola chemicko-technologická, 1986, 171 s. info
- VESPALEC, Radim. Chirální separace v kapalném prostředí. 1995, 1 svazek. info
- Assessment methods (in Czech)
- přednáška Zk
- Language of instruction
- Czech
- Further Comments
- The course is taught annually.
- Enrolment Statistics (Spring 2008, recent)
- Permalink: https://is.muni.cz/course/sci/spring2008/C8022