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Lecturer(s)
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Bednář Petr, prof. RNDr. Ph.D.
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Course content
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Instrumental aspects of high performace liquid chromatography (HPLC). Troubleshooting in HPLC - solving problems related to instrumentation, optimization of separation. Modern detection techniques in HPLC and combination of HPLC with mass spectrometry (HPLC/MS). Sample pretratment for HPLC analysis (extraction, SPE, SFE, derivatization). Capillary electrophoresis (CE) and electrochromatography (CEC), comparison with HPLC. HPLC in pharmaceutical and clinical practice. Food analysis. Analysis of (bio)polymers (GPC etc.). Separation of isomers and enantiomers (chiral discrimination). Retention and migration data measurement - utilization for identification, optimization of separation, study of interactions. Miniaturization in HPLC, capillary techniques (miniaturization in combination with mass spectrometry), CEC. Preparative chromatography. Application of HPLC to analysis of real complex samples. Discussion on particular analytical tasks.
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Learning activities and teaching methods
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Lecture, Dialogic Lecture (Discussion, Dialog, Brainstorming)
- Attendace
- 26 hours per semester
- Homework for Teaching
- 39 hours per semester
- Preparation for the Course Credit
- 55 hours per semester
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Learning outcomes
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The aim is to familiarize with practical utilization of liquid chromatography.
Application of theoretical knowledge for the solution of model problems in the field of high performance liquid chromatography
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Prerequisites
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The knowledge of basics of analytical chemistry and instrumental methods of analytical chemistry is recommended.
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Assessment methods and criteria
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Oral exam
Successful completion of the course requires active participation in at least 70% of the seminars, preparation and delivery of a short scientific presentation in MS PowerPoint on a current topic in high-performance liquid chromatography, and participation in the subsequent scientific discussion. The course is concluded with an oral colloquium conducted as a moderated group discussion covering the topics addressed during the course. The instructor directs questions to individual students, while other participants may supplement and further elaborate on their responses. Each student's performance in the colloquium is assessed individually based on independently demonstrated subject knowledge, the ability to apply this knowledge, the capacity for well-founded scientific argumentation, and active contribution to the discussion.
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Recommended literature
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Gratzfeld-Husgen A., Schuster R. HPLC for Food Analysis, A primer. Germany. 2001.
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Churáček J. Analytická separace látek. Praha. 1990.
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Churáček J., Jandera P. Úvod do vysokoúčinné kapalinové kolonové chromatografie. Praha. 1984.
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Nováková L., Douša M., Česla P. Moderní HPLC separace v teorii a praxi. I., 2. přepracované a rozšířené vydání. Brno. 2021.
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Snyder L.R., Kirkland J.J., Glajch J.L. Practical HPLC Method Development. 2nd ed. New York. 1997.
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