C7023 Separation Methods A

Faculty of Science
Autumn 2011 - acreditation

The information about the term Autumn 2011 - acreditation is not made public

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.
Department of Experimental Biology – Biology Section – Faculty of Science
Prerequisites
basic knowledge of physical and analytical chemistry is prerequisited.
Course Enrolment Limitations
The course is offered to students of any study field.
Course objectives
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.
Syllabus
  • 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 (immobilised-metal affinity chromatography); SFC (supercritical fluid chromatography); PC (perfusion chromatography), preparative chromatography – principles and carry-out
  • 9. development of chromatographic method, criteria and optimisation of separation system;
  • study and description of separation; quantitative relations between structure and selectivity in 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
Literature
  • MEYER, Veronika R. Practical High-Performance Liquid Chromatography. 3rd ed. Chichester: J. Wiley & Sons, 1999, 338 pp. 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
Teaching methods
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.
Assessment methods
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.
Language of instruction
English
Follow-Up Courses
Further comments (probably available only in Czech)
The course can also be completed outside the examination period.
The course is taught annually.
The course is taught: every week.
Listed among pre-requisites of other courses
The course is also listed under the following terms Autumn 2010 - only for the accreditation, Autumn 2008, Autumn 2009, Autumn 2010, Autumn 2011, Autumn 2012, Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020, autumn 2021, Autumn 2022, Autumn 2023, Autumn 2024.