C8022 Separation Methods B

Faculty of Science
Spring 2014
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.
National Centre for Biomolecular Research – Faculty of Science
Contact Person: doc. Mgr. Jan Havliš, Dr.
Supplier department: National Centre for Biomolecular Research – Faculty of Science
Timetable
Wed 16:00–17:50 C14/207
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
Course objectives
Separation of macromoleculs (gel permeation chromatography (GPC), light scattering, hydrodynamic chromatography (HDC), field-flow fractionation (FFF)). + Gas chromatography (GC), theory and praxis. + Separations in force field = Electromigration methods (capillary zone electrophoresis (CZE), isotachophoresis (ITP), micellar electrokinetic chromatography (MEKC), gel capillary electrophoresis (GCE)) and Mass spectrometry + Membrane separations
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 light scattering (LS), Low-angle laser light 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 (catharometer). 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 analysed by capillary electrophoresis. Proteins. Polypeptides charge. Electrophoretic titration curve. Physical principles of capillary electrophoresis - electrophoretical mobility. Electroosmotic flow - electroosmotic pump, electroosmotic flow modifiers - 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 parameters: capillary, temperature control, detectors (indirect detection), sample introduction (injection), reduction of band broadening. Optimization: heat dissipation, electroosmotic flow, on-capillary detection, molecular diffusion, band broadening due to injection. Evaluation of electrophoretic efficiency, advantages of capillary electrophoresis. Evaluation of electrophoretograms: qualitative and quantitative. Limit of detection. Validation of methods of CE. SDS - Page electrophoresis, capillary ion electrophoresis (CIE), indirect detection, chiral separation, capillary electrophoresis in non-aqueous 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
Teaching methods
the lecture is based on ppt presentation and its explication. presentation itself will be available as a study material (black-and-white printable pdf with high resolution and restricted access rights). it is recommended to attend the lecture, because of the explication, which significantly extends the presentation and because there are no available textbooks in Czech language covering certain parts of the subject.
Assessment methods
oral examination; students are required to understand and to be familiar with the principles and its applications. examination consists of three basic questions, which would be during the examination expanded to let the student demonstrate the extent of topic understanding.
Language of instruction
Czech
Further Comments
Study Materials
The course can also be completed outside the examination period.
The course is taught annually.
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 2006, Spring 2007, Spring 2008, Spring 2013, 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 2014, recent)
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