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Lecturer(s)
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Fischer Jan, doc. Ing. CSc.
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Jandera Pavel, prof. Ing. DrSc.
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Course content
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Basics of optimization of separation - quality of separation, sensitivity, amount of separation compounds, time of separation - and their influence on the optimization approach. Overview of the working parameters, which determine quality of separation. Criterions of separation quality assesment and their measuremens - resolution, Kaiser separation criterion, peak capacity, product of resolution, chromatographic optimization function. Influence of sample composition on separation, their characterisation and prediction. Basic optimisation strategy. Empiric systematic approach to optimisation of separation. One-parametric optimization of separation conditions. Window diagram method. Predictive methods of optimisation of retention and separation selectivity. Simmultaneous multiparametric optimisation - sequence and simmultaneous strategy. Simplex method. Resolution overlap area mapping. Calculation of simmultaneous optimisation. Optimisation of separation efficiency in gas and liquid chromatography. Optimisation of column dimensions, packing particles and flow of the mobile phase. Optimisation of separation temperature. Optimisation of temperature gradient and gradient elution in chromatography. Multidimensional separation techniques. Optimisation of separation conditions in capillary electrophoresis and micellar electrokinetic chromatography. Trace analysis - influence of working conditions in chromatography and electrophoresis on sensitivity and limits of detection and quantification, their optimisation. Selectivity and sensitivity of detection and their optimisation. Miniaturised separation systems - influence of miniaturisation on separation quality, practical applications and instrumental limitations. Comparison of advantages and disadvantages of capillary chromatography, capillary electrophoresis and electrochromatography. Expert systems for optimisation of chromatographic separations. Overview of commercially available systems of automatic optimisation of separation processes.
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Methods of individual activities
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Learning outcomes
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In the course the use of basic knowledge in the field of chromatographic separation and electromigration analytical techniques for the design of the working conditions for the solution of concrete problems and the separation of systematic optimization is emphasised. The aim is to obtain an overview of core strategic processes to optimize the analytical methods of separation and the relevant knowledge, so that after the completion of the course the listener is able to design optimal separation conditions for the analysis of specific samples.
Ater the completion of the course, the listener will be able to design optimal separation conditions for the analysis of specific samples.
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Prerequisites
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Course of Analytical separation methods.
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Assessment methods and criteria
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Oral examination
Seminar project, written examination.
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Recommended literature
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Churáček J. a kol. Analytická separace látek. Praha, 1990.
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Churáček J. a kol. Nové trendy v teorii a instrumentaci vybraných analytických metod. Praha, 1993.
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Snyder L. R. a kol. Practical HPLC method development. New York, 1997.
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