Electrochemical and electromigration methods

The graduates of natural sciences and life sciences area

Number of lecture hours- 6 h

Number of laboratory classes hours - 4 h

W1: knows and can describe electrokinetic phenomena in electromigration techniques

W2: knows the variants of electromigration techniques

W3: knows detection techniques in electromigration techniques

W4: knows the construction of CE apparatus

W4: knows the parameters influencing the resolution in electromigration techniques

W5: knows the methods of separation of chiral compounds

W6: has the basic knowledge of electrochemical methods of analysis

U1: can prepare the capillary and use the CE instrument

U2: can perform the CE and MEKC analysis

K2: is oriented to acquiring new knowledge

- lectures as a multimedia presentations;

- one by one and on-line consultations (distance learning);

- laboratory exercises instructions prepared based on the information given during the lectures are useful and required at the laboratory;

- individual work with the apparatus and advanced analytical equipment under the teacher supervision;

In addition, for all students hard copies are prepared as well as laboratory instructions.

The aim is to acquaint students with the theory and practice of electrochemical and electromigration analytical techniques. The special attention will be paid on electromigration methods of separation.

Lecture:

The lecture covers the basics of electrochemical analytical techniques (conductometry, ion-selective electrodes, polarography and related techniques), equipment construction and their analytical capabilities. In the second part of the lecture: electromigration techniques - electroosmosis and electrophoresis, variants of electromigration methods such as capillary zone electrophoresis (CZE), izotachophoresis (ITP), capillary gel electrophoresis (CGE), micellar electrokinetic chromatography (MEKC), elektrochromatography (CEC) and capillary isoelectric focusing (CIEF) will be discussed. The apparatus construction, operation, analytical capabilities and example applications, chip-type separation systems will be presented as well.

Lecture:

Electromigration techniques - lecture includes discussion on principles that governs the separation conditions in capillary electrophoresis, in its various variants; discussion on the electrophoretic techniques variants, their differences in how to perform the separation, detection methods and the presentation of their advantages, disadvantages and limitations; discussion about the applications and chips techniques.

Separation techniques in supercritical conditions - lecture include: the definition and properties of supercritical fluids, provide the benefits of supercritical mobile phase, the construction of typical SFC chromatographs, used detectors, columns used in SFC, methodology to carry out the separation with special attention paid on the possible use of pressure gradients, temperature, mobile phase composition, a discussion of typical applications and comparisons with HPLC and GC.

Separation of optically active compounds - lecture include: chirality definition, a review of the exemplary chiral separation performed by GC, HPLC, CZE, MEKC, CEC; choice and examples of chiral selectors, direct and indirect methods; derivatization.

Extraction at elevated pressure and temperature (SFE and ASE) - lecture covers: the definition of supercritical fluids, provide the benefits of supercritical fluid as a extraction medium, the aims and methods of modifiers use, SFE apparatus design and principles; restrictors and their types – differences in principle of work, manufacture methods, their advantages and disadvantages, selection of extraction conditions, applications, the principle of ASE extraction, the benefits usage liquid under high pressure and high temperature, construction of ASE apparatus and its operation, ASE process steps, limitations, advantages and disadvantages of ASE methods, exemplary applications and comparison with other extraction techniques.

Application of gas chromatography in food analysis, pharmacy, environmental protection, etc. – lecture presents the typical and less known applications of gas chromatography, fast gas chromatography (Fast GC) - the method principle, the columns and apparatus construction and work, two-dimensional chromatography;

Electrochemistry in ecoanalytics - lecture covers the basics of electrochemical methods: polarography, amperometry, conductometry, potentiometry, coulometry, the application of these techniques as a detection systems in electromigration techniques and chromatography will be also discussed.

Laboratory classes:

Optimization of separation conditions in micro and electromigration techniques: micro-HPLC chromatograph construction, the special focus on the influence of extracolumn volume on the separation system efficiency; the methods of preparation of chromatography columns, set-up for the capillary and classical columns packing - display; preparation of fused silica capillaries for capillary electrophoresis, the detection window preparation (practical works), construction of CE apparatus, the comparison of the vitamin mixture separation by CE and MEKC - discussion on the separation mechanism.

Supercritical fluid extraction (SFE), P & T, SDE, ASE - discussion on the construction and operation of SFE and ASE apparatus, preparation of the sample for extraction, selection of the extraction conditions - caffeine extraction from coffee or tea with the final determination by MEKC - an example.

Optimization of the separation process in GC I (GC construction, temperature gradient, flow optimization, Fisons) - discussion about the gas chromatograph construction, preparation of the GC for analysis, determination of the optimum carrier gas flow, BTEX mixture separation in isocratic and gradient conditions.

Analysis of pharmaceuticals and natural products coupled with separation techniques: SFE/ASE - LC/MSn, LC/MS.

During the laboratory classes students also become familiar with:

1. Equipment construction;

2. Methodology of preparation of capillary for electromigration techniques;

3. Column quality evaluation;

4. Principles of separation under CZE and MEKC conditions. Separation mechanisms also will be discussed.

1. D. Baker, Capillary electrophoresis, Wiley & Sons, INC, New York, 1995

2. A. Cygański, Podstawy metod elektroanalitycznych, Warszawa, Wydawnictwa Naukowo-Techniczne, 2004.

3. Z. Brzózka, (red.) Miniaturyzacja w analityce, Warszawa, Oficyna Wydawnicza Politechniki Warszawskiej, 2005.

4. Bernd Wenclawiak (red.) Analysis with supercritical fluids: Extraction and Chrmatography, Springer-Verlag, 1992.

5. K. Oukebdane, F. Portet-Koltalo, N. Machour, F. Dionnet, P.L. Desbène Comparison of hot Soxhlet and accelerated solvent extractions with microwave and supercritical fluid extractions for the determination of polycyclic aromatic hydrocarbons and nitrated derivatives strongly adsorbed on soot collected inside a diesel particulate filter, Talanta, 82 (2010) 227-236

6. M. Lammerhofer Chiral recognition by enantioselective liquid chromatography:Mechanisms and modern chiral stationary phases, J. Chromatogr. A 1217 (2010) 814-856

Student - graduate student:

1. identifies and distinguish between issues related to the classes topic;

2. are able to find and use the required literature in English and Polish;

3. are using, applying and explaining the subject terminology related

to the study issues (not using laboratory slang) in English

and its counterparts in Polish;

4. are applying in practice the theoretical knowledge how to operate

by the apparatus and small laboratory equipment in a correct and proper way as well. Performing measurements independently based on the knowledge he/she gained;

5. preparing samples for analysis by him-/herself, creating analytical procedures and standard procedures;

6. independently examines, interprets and calculates the results obtained

in the laboratory. Applying the appropriate analytical procedures. Preparing writing research reports and notes, which may be the useful for issuing the scientific publications;

7. compares, explains and describes obtained results in comparison with the available standards and literature. Predicting behavior and probable scenarios in the laboratory during sample preparation as well as the same study (e.g. principles of health and safety in the workplace).

The pass mark class is attendance at lectures and active participation, knowledge of analytical procedures is required for proper implementation

of the exercise-laboratory analysis.

Course assessment is based on the frequency and participation in the classes as well as receiving a positive evaluation at an oral, final exam.

not provided