To learn the principles of analytical techniques and their applications in the chemical, biological and food fields.
Knowledge of basic principles of general and analytical chemistry
Course contents summary
Principles of chromatography and capillary electrophoresis. Coupling to mass spectrometry.
Applications: PROTEOMICS, FOODS, CLINICALS
Sampling and preparation of the analytical sample. Chromatography: selection criteria of the cormatographic parameters for the optimization of the resolution. Gas chromatography: Principles and instrumentation. Columns and stationary phases. High performance liquid chromatography (HPLC): principles and instrumentation. Features of fixed and mobile phases. Ionic Chromatography (IC): principles and instrumentation. Specific detectors for ionic chromatography: conductimetric and amperometric. Application of strong anionic exchange chromatography with pulsed amperometric detection (HPAEC-PAD) in the separation and characterization of oligo and polysaccharides. Features of fixed and mobile phases. Dimensional exclusion chromatography. Elettroseparazioni. Principles. Gel electrophoresis. Capillary electrophoresis: theory and applications. Capillary micellar electrocinetic chromatography. Fundamentals of mass spectrometry, main ionization techniques. Ion sources; hard and soft ionization techniques in GC-MS: electronic ionization (EI) and chemical ionization (CI). Ionization techniques in LC-MS: chemical ionization at atmospheric pressure (APCI), atmospheric pressure photoionization (APPI), ionization by electronbulization (ESI). CE-MS coupling. Source for laser-assisted laser desorption / ionization (MALDI) and ionization -SELDI (Surface Enhanced Laser Desorption / ionization). Mass analyzers: magnetic field and electro-magnetic (double focusing) quadrupole, ion trap, flight time (TOF). Fourier transform mass spectrometry: Orbitrap and cyclotonic ion resonance analyzer. Hybrid instruments. Detectors and data acquisition. Tandem mass spectrometry. the interpretation of mass spectra; procedures for interpreting mass spectra of organic molecules obtained by electron ionization and other ionization techniques. Applications and quantitative analysis. Mass spectrometry coupling with chromatographic, gaseous and liquid and electrophoretic capillary techniques. Application of mass spectrometry techniques to the study of biomolecules: examples of development of GC-MS, LC-MS and CE-ME methods. Proteomics; Botton-up and Top-down approaches.