Course: Derivatization for Separation Techniques

« Back
Course title Derivatization for Separation Techniques
Course code KALCH/C568
Organizational form of instruction Lecture
Level of course Master
Year of study 1
Semester Summer
Number of ECTS credits 2
Language of instruction Czech
Status of course Compulsory-optional
Form of instruction Face-to-face
Work placements This is not an internship
Recommended optional programme components None
Lecturer(s)
  • Česla Petr, doc. Ing. Ph.D.
  • Kašpar Michal, Ing. Ph.D.
Course content
Utilization of chemical reactions in separation analytical techniques - definition of terms, basic principles and procedures, classification of derivatization reactions. Apparatures for derivatization, samples and reaction vessels, heating and evaporation, manipulation with a sample and reactants, removal of derivatization agents, serial derivatizations using robotic systems. Principles of derivatizations for gas chromatography - influence of derivatives on retention characteristics, types of reactions, derivatization conditions, instrumental arrangement. Application of chemical reactions for derivatization in gas chromatography - silylation reactions, esterification, alkylation, acylation, preparation of cyclic derivatives. Hydrogenation, dehydrogenation, ozonolysis, hydrolytic methods. Pyrolysis gas chromatography - principles, development of pyrolysis methos, commercial pyrolyzers, automatic systems, applications. Derivatization for liquid chromatography - types of derivatization reactions, precolumn derivatization. Postcolumn derivatization, influence of reactor on efficiency of separation, construction of flow-through reactors, reactors with segmented flow of mobile phase. Utilization of chemical reactions for derivatization in liquid chromatography with UV detection - the most common chromophores and fluorophores, esterification, reaction with benzoylation agents, reactions with iso- and isothiocyanates. Derivatization for fluorescent detection - principles, agents, reaction conditions. Preparation of derivatives for electrochemical detection and less common types of detectors. Derivatization for capillary electrophoresis - techniques enhancing sensitivity of detection, on-line techniques of derivatization in separation capillary, introducing charge to a molecule, increasing hydrophobicity, separation of enantiomers. Derivatization for separation techniques coupled with mass spectrometry - ionization of analytes, fragmentation, structural analysis, application of isotopically labeled chemicals.

Learning activities and teaching methods
Monologic (reading, lecture, briefing)
Learning outcomes
The aim of the course is the introduction of application of chemical reactions in analytical separations. Students wll be introduced to theoretical background of derivatization, practical methods and applications in common separation techniques used in analytical laboratories.
Graduate of the course is able to assess the usage of different derivatization procedures and methods and critically evaluate achieved results. Graduate is aware of the fundamentals, instrumentation and the use of various derivatization reactions for chromatographic and electromigration separation techniques.
Prerequisites
Basic courses of Instumental Analytical Chemistry, Physical Chemistry and Organic Chemistry.

Assessment methods and criteria
Student performance assessment

Written examination, consisting theoretical questions and practical examples of derivatization agents and reactions
Recommended literature
  • Blau K., Barbre A. Handbook of Derivatives for Chromatography, Second Edition, 1993, John Wiley & Sons, Chichester.
  • Komárek, Karel. Reakční chromatografie v organické analýze. Praha: Státní nakladatelství technické literatury, 1989. ISBN 80-03-00153-6.
  • Lunn, George. Handbook of derivatization reactions for HPLC. New York: John Wiley & Sons, 1998. ISBN 0-471-23888-0.
  • Wilson, Ian D. Handbook of methods and instrumentation in separation science. London: Elsevier , 2009. ISBN 978-0-12-375095-2.


Study plans that include the course
Faculty Study plan (Version) Category of Branch/Specialization Recommended year of study Recommended semester