C8024 Separation methods B

Přírodovědecká fakulta
jaro 2024
Rozsah
2/0/0. 2 kr. (příf plus uk plus > 4). Doporučované ukončení: zk. Jiná možná ukončení: k.
Vyučující
doc. Mgr. Jan Havliš, Dr. (přednášející)
Garance
doc. Mgr. Jan Havliš, Dr.
Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Dodavatelské pracoviště: Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Předpoklady
C7023 Separation methods A
basic knowledge of physical and analytical chemistry + subject C7023 separation methods A are pre-requisited.
Omezení zápisu do předmětu
Předmět je otevřen studentům libovolného oboru.
Cíle předmětu
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).
Výstupy z učení
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.
Osnova
  • 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
Literatura
  • Capillary electrophoresis : methods and protocols. Edited by Philippe Schmitt-Kopplin. Totowa, N.J.: Humana Press, 2008, xix, 809. ISBN 9781588295392. info
  • Analysis and detection by capillary electrophoresis. Edited by M. L. Marina - M. Valcárcel Cases - Ángel Ríos Castro. 1st ed. Amsterdam: Elsevier, 2005, xxviii, 76. ISBN 0444517189. info
  • PLEVA, Michael A. a Frank A. SETTLE. Gas chromatography - mass spectrometry a knowledge base. Amsterdam: Elsevier, 1988, 12 s. ISBN 0444427619. info
  • AMBROSE, D. Gas chromatography. 2nd ed. London: Butterworths, 1971, 321 s. ISBN 0-408-70129-3. info
Výukové metody
the lecture is based on ppt presentation and its explication. presentation it-self 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 of limited availability of textbooks in english covering certain parts of the subject. subject will be eventually lectured if the number of enrolled students exceeds number 5, otherwise students will be given the chance to pass the subject without lecturing combining study materials and on-demand tutorial (at max 2x2h per semester; contact the lecturer to set up the schedule).
Metody hodnocení
written examination; students are required to understand and be familiar with the principles and their applications. examination consists of three basic questions.
Vyučovací jazyk
Angličtina
Další komentáře
Studijní materiály
Předmět je vyučován každoročně.
Výuka probíhá každý týden.
Předmět je zařazen také v obdobích jaro 2011 - akreditace, jaro 2009, jaro 2010, jaro 2011, jaro 2012, jaro 2012 - akreditace, jaro 2013, jaro 2014, jaro 2015, jaro 2016, jaro 2017, jaro 2018, jaro 2019, jaro 2020, jaro 2021, jaro 2022, jaro 2023, jaro 2025.