C8022 Separation Methods

Faculty of Science
Spring 2006
Extent and Intensity
2/0/0. 2 credit(s) (fasci plus compl plus > 4). Recommended Type of Completion: zk (examination). Other types of completion: k (colloquium).
Teacher(s)
doc. Mgr. Jan Havliš, Dr. (lecturer)
doc. RNDr. Bc. Jiří Pazourek, Ph.D. (lecturer)
Guaranteed by
doc. RNDr. Bc. Jiří Pazourek, Ph.D.
Department of Chemistry – Chemistry Section – Faculty of Science
Timetable
Mon 11:00–12:50 02004
Prerequisites
C7021 Separation Methods A
C3100 + C4050 Analytical chemistry (at least C1660) C7021 Separation methods
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
Course objectives
Liquid chromatography (LC), theory and praxis. Separation of macromoleculs (gel permeation chromatography (GPC), light scattering, hydrodynamic chromatography (HDC), field-flow frakcionation (FFF)) + elektroforetic methods = capillary zone electrophoresis (CZE), izotachophoresis (ITP), micelat electrokinetic chromatography (MEKC), gel capillary electrophoresis (GCE)
Syllabus
  • Liquid-liquid extraction. Organic molecules extraction, influence of coexisting phases on extraction, the relationship between analyte structure and its partition coefficient. Extraction of chelates, extraction of ion associates, extraction solvents, influence of pH, synergism, applications of analytical reactions. Extraction flow analysis (FIA), multiple extraction systems, continual extractions, chromatography. Principle of chromatographic separatrion, relation to extraction, theoretical plate, resolution, efficiency, selectivity and retention. Basics of separation optimization, High pressure liquid chromatography (HPLC). Classification of basic separation mechanisms in liquid chromatography (liquid-liquid chromatography, LLC, liquid-solid chromatography, LSC, ion chromatography, IC, gel permeation chromatography, GPC) and their possible combination (ion exclusion in IC, adsorption in GPC etc.) Criteria of selection and optimalization of mobile phases, LC column packing, reverse phase liquid chromatography. Instrumentation of liquid chromatography, pumps, injectors, detectors (diode array detectors, DAD), fluorescence, amperometric, conductivity, refrectometric detectors, miniaturization in LC, applications. 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. Electrophoretic 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.
Teacher's information
http://www.chemi.muni.cz/~pazourek/student/sepmetody.html
The course is also listed under the following terms Spring 2008 - for the purpose of the accreditation, Spring 2000, Spring 2001, Spring 2002, Spring 2003, Spring 2004, Spring 2005, Spring 2007, Spring 2008, Spring 2013, Spring 2014, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2022, Spring 2023, Spring 2024, Spring 2025.
  • Enrolment Statistics (Spring 2006, recent)
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