C8022 Separation Methods B

Faculty of Science
Spring 2025
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).
In-person direct teaching
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
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
main objective of the course is to acquaint students with separation methods based on analyte size, separation in force fields and on membranes; at the end part also with other aspects of analytical separations (chiral separation, optimisation, validation).
Learning outcomes
at the end of the course students should be able:
to master principles of macromolecular separation, mass spectrometry, electromigration separation and memebrane separation
to understand the wide scope of their contemporary implementation and utilisation
to optimisation of separation system, to evaluate its efficiency and to understand basic procedures of method validation.
Syllabus
  • 1. separation of macromolecules: history, theoretical principles – macromolecule description, molecular mass; modes of macromolecular separation – SEC (size exclusion chromatography), carry-out of SEC – columns,
  • 2. detectors (spectrophotometry, viscosimetry, osmometry, light scattering), HDC (hydrodynamic chromatography), FFF (field-flow fractionation) – principles, instrumentation, modes (SdFFF, ThFFF, EFFF, FFFF).
  • 3. electromigration methods: history, theoretical principles – separation in outer field, separation description (ion mobility, EOF electroosmotic flow), efficiency and factors, which influence it (heat, diffusion, dispersion); arrangement of electromigration methods – injection, voltage, separation environment (capillary, membrane, gel, chip).
  • 4. detectors (spectrophotometric, laser induced fluorescence, conductivity, mass spectrometry etc.); preparative modes; electrophoretic system definition and analytical information in electrophoretogram; basic modes of electromigration methods – capillary zone electrophoresis (CZE), capillary gel electrophoresis (CGE), planar gel electrophoresis (GE), capillary isoelectric focusing (CIEF), capillary isotachophoresis (CITF), micellar electrokinetic chromatography, thin-layer electrochromatography, capillary electrochromatography;
  • 5. affinity and non-aqueous electrophoresis; mass spectrometry (as a separation method): history, theoretical principles – separation in outer field, mass spectrum, basic terms; ionisation (collision, chemical reaction, electrospray), 6. mass analysis (electric sector, magnetic sector, quadrupole, ion trap, time-of-flight analyser, orbital trap, drift cell, ion cyclotron resonance), tandem mass analysis, hybride mass analysers, ion detection (multiplier, induction), vacuum technique;
  • 7. membrane separation: history, theoretical principles – non-equilibrium separation, membrane separation principles, carry-out (batch, continual), membranes (selectivity, materials, fouling), separation space – modules: membrane, spiral-wound, tubular, hollow-fibre, rotation, basic modes of membrane separations: microfiltration, ultrafiltration, nanofiltration, reversed osmosis, electrodialysis, pervaporation, membrane distillation, gas permeation, dialysis.
  • 8. chiral separation: definition of chirality; optical rotation dispersity; principals of chiral separation – chiral selectors, chiral separation methods including preparative, chiral selectors free, immobilised and derivatization.
  • 9. development of chromatographic method, criteria and optimisation of separation system; single and multiple criterial of separation quality evaluation; validation of analytical separation method; why and how, elemental terms –method identity, method precision, calibration, selectivity, robustness
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
The course can also be completed outside the examination period.
The course is taught annually.
The course is taught: every week.
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 2014, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2022, Spring 2023, Spring 2024.
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