E0210 Chromatographic Methods

Faculty of Science
Autumn 2020
Extent and Intensity
2/0/0. 2 credit(s) (plus extra credits for completion). Type of Completion: zk (examination).
Teacher(s)
PharmDr. Zdeněk Spáčil, Ph.D. (lecturer)
Guaranteed by
PharmDr. Zdeněk Spáčil, Ph.D.
RECETOX – Faculty of Science
Contact Person: PharmDr. Zdeněk Spáčil, Ph.D.
Supplier department: RECETOX – Faculty of Science
Timetable
Tue 11:00–12:50 D29/252-RCX1
Prerequisites
The course requires a basic knowledge of analytical chemistry, the insight into instrumental separation methods is an advantage.
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
fields of study / plans the course is directly associated with
Course objectives
The main objective of the course is to explain the basic theory of chromatographic separation applicable to modern chromatographic techniques. Students will acquire expertize necessary to choose an appropriate instrumentation for given analysis and to optimize experimental conditions of chromatographic methods, which are applicable in many research areas as well as in private companies.
Learning outcomes
Students completing the course will be able to:

- understand basic principle of the most commonly used; chromatographic methods and some other analytical separation techniques;

- know important practical and technical aspects of modern chromatography instrumentation applicable and highly relevant to scientific work;

- propose a protocol and technical implementation of chromatographic methods in relation to the type of analysis and the physico-chemical properties of the analyte;

- explain generic separation theory of chromatographic separation and underlying mechanisms and select suitable chromatographic system, stationary phase and separation mode to obtain high-quality scientific data;

- apply chromatographic theory to characterize analytical systems, surface and material of stationary phase, and supposed interaction of the analyte within the system;

- evaluate and interpret results of chromatographic analysis;

- assess the practical relevance of the combination of chromatographic techniques with different types of detection techniques, in particular with spectroscopic and spectrometric techniques;

- overview current trends in the instrumentation, stationary phases and method development - utilize chromatographs for qualitative and quantitative determination of various analytes in biological matrices;

- select and optimize experimental conditions of separation techniques suitable to increase selectivity, separation efficiency, number of identifications and optimal quantification limits for the most sensitive analysis
Syllabus
  • 1. Basic terms of chromatographic separation, retention parameters, chromatogram, chromatographic resolution, separation efficiency, broadening of elution zone, Knox and van Deemterova equation.
  • 2. Liquid Chromatography (LC), column and planar chromatography, chemical equilibrium, theoretical floor, peak capacity, separation conditions.
  • 3. Molecular basis of migration, longitudinal diffusion, sorption and desorption, mass transfer, flow diffusion.
  • 4. Chromatographic column, types of stationary phases, chemistry and chromatographic properties of sorbents.
  • 5. Mobile phases, classification of solvents, buffers and common additives, mobile phase for gradient elution, optimization techniques, elution modes.
  • 6. Instrumentation for high performance (HPLC) and ultra-high performance (UHPLC) liquid chromatography, mobile phase transport, sample injection, detector types and principle.
  • 7. Chromatography: normal-phase (NP), reversed-phase (RP), ion-exchange (IEC), supercritical fluid (SEC), gel permeation (GPC).
  • 8. Gas chromatography.
  • 9. Mobile phases and stationary phases for GC, comparison of gas properties.
  • 10. GC instrumentation, sample injection, detector types and principle.
  • 11. Trends in chromatography, development of stationary phases, derivatization of analytes, micro- and nano-flow chromatography.
  • 12. Examples of application of chromatographic methods in various research areas.
Literature
    recommended literature
  • GIDDINGS, John Calvin. Unified Separation Science. Wiley, 1991, 352 pp. ISBN 0-471-52089-6. info
  • ROBARDS, Kevin, Paul R. HADDAD and Peter E. JACKSON. Principles and practice of modern chromatographic methods. London: Academic Press, 1994, x, 495. ISBN 0125895704. info
  • NOVÁKOVÁ, Lucie and Michal DOUŠA. Moderní HPLC separace v teorii a praxi. 1. vyd. [Klatovy]: Michal Douša, 2013, 299 s. ISBN 9788026042433. info
  • NOVÁKOVÁ, Lucie and Michal DOUŠA. Moderní HPLC separace v teorii a praxi. 1. vyd. [Klatovy]: Michal Douša, 2013, 235 s. ISBN 9788026042440. info
  • SPÁČIL, Zdeněk. Aplikace kapalinové chromatografie za ultravysokého tlaku a hmotnostní spektrometrie v analýze polyfenolických látek. Hradec Králové, 2010. Dizertační práce. Univerzita Karlova, Farmaceutická fakulta v Hradci Králové.
Teaching methods
The lessons are performed in the form of PowerPoint presentation. Prior each lecture, students will receive printed materials for writing their own notes and queries. The comprehensibility of the difficult parts is tested interactively.
Assessment methods
The presence at the lecture is not compulsory, but highly recommended for facile and fluent understanding of lectured topics. The emphasis is placed on interaction with students to encourage discussion on the matter. Newly acquired knowledge is verified by an oral exam.
Language of instruction
Czech
Follow-Up Courses
Further comments (probably available only in Czech)
Study Materials
The course can also be completed outside the examination period.
The course is taught annually.
Teacher's information
https://is.muni.cz/auth/predmet/sci/akreditace2017podzim/E0210
The course is also listed under the following terms Autumn 2019, autumn 2021, Autumn 2022, Autumn 2023, Autumn 2024.
  • Enrolment Statistics (Autumn 2020, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2020/E0210