PřF:C8024 Separation Methods B - Course Information

C8024 Separation Methods B

Extent and Intensity

2/0/0. 2 credit(s) (fasci plus compl plus > 4). 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
Supplier department: National Centre for Biomolecular Research – Faculty of Science

Prerequisites

C7023 Separation Methods A
basic knowledge of physical and analytical chemistry + subject C7023 separation methods A are pre-requisited.

Course Enrolment Limitations

The course is offered to students of any study field.

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

• Capillary electrophoresis : methods and protocols. Edited by Philippe Schmitt-Kopplin. Totowa, N.J.: Humana Press. xix, 809. ISBN 9781588295392. 2008. info
• Analysis and detection by capillary electrophoresis. Edited by M. L. Marina - M. Valcárcel Cases - Ángel Ríos Castro. 1st ed. Amsterdam: Elsevier. xxviii, 76. ISBN 0444517189. 2005. info
• PLEVA, Michael A. and Frank A. SETTLE. Gas chromatography - mass spectrometry a knowledge base. Amsterdam: Elsevier. 12 s. ISBN 0444427619. 1988. info
• AMBROSE, D. Gas chromatography. 2nd ed. London: Butterworths. 321 s. ISBN 0-408-70129-3. 1971. 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 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).

Assessment methods

written examination; students are required to understand and be familiar with the principles and their applications. examination consists of three basic questions.

Language of instruction

English

Further Comments

Study Materials
The course is taught annually.
The course is taught: every week.

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