Syllabus
* successful passing of lectures C230P05N: Separation methods or C230P51: Separation methods (kata) is required before entering this course
1.Introduction to HPLC, its importance and practical application, comparison to other separation methods. General scheme and pieces of HPLC equipment. Basic terms and definitions. Types of analytes' interactions, separation systems.
2. Kinetic and thermodynamic aspects of separation in HPLC. Distribution constant, retention factor, performance of chromatographic column, band broadening, peak shapes, chromatographic resolution.
3. Separation systems: Normal-phase and reversed-phase chromatography.
4. Separation systems: Ion-exchange chromatography. Affinity chromatography. Size-exclusion chromatography..
5. Instrumentation: Pumps, injectors, columns, flow splitters, fractions' collectors. Preparative HPLC.
6. Detection: Principles of detection in HPLC; detectors, their construction and parameters.
7. Qualitative and quantitative analysis in HPLC.
8. Practical aspects of HPLC : Samples' and mobile phases' preparation, operation and service of HPLC instrument, problem solving. Commercially available HPLC equipments.
9. Aplications of HPLC in pharmaceutical analysis, food and beverage analysis, environmental analysis.
10. Aplication of HPLC in analysis of bio/macromolecules.
11. Modern trends in HPLC. Multidimensional separation systems. Miniaturization, microfluid systems. New types of stationary phases and their properties.
12. Excursion to routine commercial HPLC laboratory - Quinta Analytica company.
Annotation
The lecture focuses on high-performance liquid chromatography (HPLC), one of the most frequently used separation technique in current analytical chemistry. The students will learn the theoretical fundamentals and get a comprehensive picture about practical usage of the method.
The lecture series is prefaced by basic terms, retention characteristics and mechanisms taking effects. A considerable attention is paid to the instrumentation, particularly to the description of individual components of the HPLC systems.
The students will learn about development of HPLC in recent years and the newest trends in the field. The next lectures are focused on HPLC daily practice and an overview about the main application areas is given.
The lecture series is closed by an excursion to routine HPLC laboratory.