C7023 Separation methods A

Přírodovědecká fakulta
podzim 2013
Rozsah
2/0/0. 2 kr. (plus ukončení). 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
Rozvrh
Po 15:00–16:50 C02/211
Předpoklady
basic knowledge of physical and analytical chemistry is prerequisited.
Omezení zápisu do předmětu
Předmět je otevřen studentům libovolného oboru.
Cíle předmětu
main objectives of lecture: introduction into liquid chromatography as analytical separation method; mastering its principles; acquainting with the wide scope of its contemporary implementation and utilisation; definition of terms pre-analytical separation, separations analytical and preparative; choice and optimisation of separation system, evaluation of its efficiency, with validation of separation method and evaluation of the separation results.
Osnova
  • 1. introduction; extraction: description of extraction equilibria in system liquid-liquid and in system liquid-solid; choice of extraction system, carry-out of extraction, influence of analyte-solvent interaction, repeated extraction
  • 2. preanalytical sample preparation: SFE (supercritical fluid extraction); ASE (accelerated solvent extraction); MAE (microwave assisted extraction), TLC (thin-layer chromatography, HPTLC 2D TLC),
  • 3. SPE (solid phase extraction); SPME (solid phase microextraction), HSE (head-space extraction)
  • analytical separation; liquid chromatography (LC) – history, theoretical ground; classification of basic separation mechanisms in LC (liquid-liquid chromatography, LLC, liquid-solid chromatography, LSC, ion chromatography, IC, gel permeation chromatography, GPC),
  • 4. physico-chemical description of LC processes, mass transfer in LC, thermodynamic aspects of separation; kinetic aspects of separation,
  • 5. LC arrangement: mobile phase delivery, elution and elution force (isocratic and gradient); injection devices; separation column, types of stationary phases (particle, monolithic, on-chip), overview of sorbents;
  • 6. detectors (diode array detectors, refractometric, fluorescence, amperometric, conductivity, light-scattering, mass spectrometric) description and evaluation of separation results: defining the chromatographic system, analytical information in chromatogram (qualitative, quantitative); precision, accuracy, limits of analysis (MDL, LOD, LOQ); evaluation of separation efficiency, testing of the system and its “good” behaviour;
  • basic LC modes: NP-HPLC (normal phase liquid chromatography), RP-HPLC (reverse phase liquid chromatography), ultra-high performance chromatography, high temperature liquid chromatography, elevated-temperature ultra-high performance chromatography,
  • 7. hydrophilic interaction chromatography (HILIC), hydrophobic interaction chromatography (HIC) IEC (ion exchange chromatography) – principles and carry-out, chromatofocusation,
  • 8. affinity chromatography – principles, IMAC (immobilized-metal affinity chromatography); SFC (supercritical fluid chromatography); PC (perfusion chromatography), preparative chromatography – principles and carry-out
  • 9. development of chromatographic method, criteria and optimization of separation system;
  • study and description of separation; quantitative relations between structure and selectivity with regard to retention;
  • 10. chiral separation: definition of chirality; optical rotation dispersity; chiral selectors and their application in separation methods (HPLC)
  • 11. validation of analytical separation method; why and how, elemental terms – method identity, method precision, calibration, selectivity, robustness
Literatura
  • MEYER, Veronika R. Practical High-Performance Liquid Chromatography. 3. vyd. Chichester: J. Wiley & Sons, 1999, 338 s. ISBN 0-471-98372-1. info
  • A practical approach to chiral separations by liquid chromatography. Edited by Ganapathy Subramanian. Weinheim: VCH Verlagsgesellschaft, 1994, 405 s. ISBN 3-527-28288-2. 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.
Metody hodnocení
oral examination; students are required to understand and 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.
Vyučovací jazyk
Angličtina
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