Learning objectives
The course, through theoretical lessons and theoretical and practical exercises, also in laboratory, will give the students an applicative knowledge on the use of the chromatographic techniques in organic chemistry.
Prerequisites
In order to proficiently attend the course, a basic knowledge of Organic Chemistry and Instrumental Analytical Chemistry is strongly suggested.
Course unit content
Correlations between chromatographic parameters (capacity factor, selectivity, theoretical plate numbers, resolution) and physico-chemical parameters of the partition process, application to the development of chromatographic methods.
GC column types, chemistry of the stationary phases, eluotropic series of liquid stationary phases, intermolecular interactions analyte-stationary phase, criteria for the choice of the stationary phases, elution orders, column parameters, elution temperature and influence on the chromatographic parameters, isothermal and gradient elution. GC detectors: applicability to organic molecules and criteria for the choice, GC/MS. Derivatization reaction of organic molecules for GC analysis: silylations, alkylations, acylations. Derivatization of triglycerides, amino acids, carbohydrates.
Thin layer chromatography (TLC), calculation of the chromatographic parameters in TLC and their physico-chemical significance, separation mechanisms, stationary phases, eluents, strenght and selectivity, Snyder classification, detection methods in TLC, flash chromatography.
High Pressure Liquid Chromatography, chemical composition of the stationary phases, normal pahse and reverse phase, separation mechanisms, polarity scales for the eluents, ionic exchange HPLC, ion pairing reverse phase HPLC, ion suppression reverse phase HPLC, hydrophobic interaction and size exclusion chromatography, micro- and nanoHPLC, UPLC. Use of Uv-vis and fluorescence detectors in correlation with the physico-chemical features of the molecules. HPLC/MS hyphenation and detection modes. Preparative HPLC with scale-up methods. HPLC applied to the determination of amino acids, peptides, proteins, carbohydrates and natural organic substances.
Chiral separations. Direct and indirect resolution, chiral stationary phases, chiral mobile phases.
Several practical lessons will be held on the instruments by developing GC, TLC, low pressure chromatography and HPLC methods in several applications.
Bibliography
Brown, Foote. 'Chimica Organica' Ed. Edises
High Performance Liquid Chromatography. ACOL, John Wiley and Sons
Gas Chromatography. ACOL. John Wiley and Sons
Scientific literature found on specialized chemical journals
Teaching methods
Frontal lectures and laboratory practice
Assessment methods and criteria
Written and oral examination
