Learning outcomes of the course unit
Knowledge and understanding
Understanding regarding the principles of analytical techniques and their applications in the chemical, biological and food fields.
knowledge regarding the instrumentation, ionization systems, ion separation and hyphenation with the separation techniques used in mass spectrometry. - Acquire the basic knowledge for the interpretation of a mass spectrum obtained in an electronic impact. Comparison with other ionization techniques such as chemical ionization
- knowledge of the potentials, limits and problems related to the type of ionization, analyzer and data acquisition method.
- knowledge regarding the chromatographic and electrophoretic parameters useful for the use of the best instrumental analytical technique to solve an analytical problem, including sample preparation techniques.
The student have to be able to assess the potential and critical points of the various analytical techniques and methodologies as well as make the correct decisions on how to deal with a problem related to the type of the sample under consideration and evaluation of the analytical data obtained.
The student have to be able to demonstrate the necessary capacity in describing with the appropriate and more pertinent technical terms.
The student must demonstrate to be able to use independently and critically the teaching material provided by the teacher, as well as other independently found , demonstrating the degree of scientific growth gained.
Knowledge of basic principles of general and analytical chemistry
Course contents summary
Principles of chromatography and capillary electrophoresis. Coupling to mass spectrometry.
Ion sources. Electronic ionization; chemical Ionization; APCI: APPI; ESI; MALDI
Mass Analysers: Tandem 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.
Lecture notes and power point presentations provided by the teacher. All the material presented in class is available online
Classroom exercises (interpretation of mass spectra, simulation optimization of separative methods of ifenated systems (GC-MS; LC-MS, CE-MS).
Laboratory (GC-MS, LC-MS).
Assessment methods and criteria