Industrial pharmacy REPLICA LATINA

High-performance chromatographic techniques. Theory and principles. Main application fields of chromatography. High Performance Liquid Chromatography (HPLC). General aspects. Chromatographic parameters (capacity factors, selectivity, efficiency, resolution). Van Deemter equation. Chromatographic supports and their physico-chemical properties: surface chemistry, specific surface area, pore size and volume. Bonded-phases: chemistry and stability. Separation mechanisms: liquid-liquid, liquid-solid, normal-phase (NP), reversed-phase (RP). Ion-Exchange Chromatography (IEC). Paired-Ion Chromatography (PIC). Size-Exclusion Chromatography (SEC). Hydrophobic-Interaction Chromatography (HIC), Hydrophilic interaction Chromatography (HILIC). Elution mode: isocratic and gradient. Detectors. UHPLC: General aspects. UHPLC stationary phases. High-Resolution Gas Chromatography (HRGC). General aspects. Microbore and packed microcapillary columns. Stereoselective and enantioselective separations. Direct and indirect approach. Main interactions between selector and selectands. Thermodynamics of enantiomers separation. Chiral stationary phases (CSPs) for HPLC and HRGC. Infrared Spectroscopy (IR) General aspects. Theory of Infrared Spectroscopy (IR). Main functional groups and typical IR absorptions of organic molecules. Basics of Fourier Transform IR (FT-IR) spectrum interpretation. Identification and structure elucidation of organic molecules by interpretation of FT-IR spectra. Ultraviolet Spectroscopy (UV) General aspects. Theory of Ultraviolet Spectroscopy (UV). Lambert-Beer law. Electronic transitions σ→σ*, n→σ*, n→π*, π→π*. UV instrumentation and solvents. Typical UV absorbing of compounds featuring only σ bonds, of aliphatic compounds with n electrons and of compounds with π electrons. Aromatic systems. Nuclear Magnetic Resonance (NMR) General aspects. Theory of Nuclear Magnetic Resonance (NMR). Nuclear magnetic moment, spin number, angular momentum, magnetogyric ratio. Larmor precession. Relaxation processes: spin-lattice and spin-spin. Proton Magnetic Resonance Spectrometry ( 1H-NMR). Important concepts and parameters in NMR. Instrumentation and sample handling. Chemical shift (diamagnetic anisotropy, bond anisotropic effect, ring-current effect, hydrogen bond, solvent polarity and magnetic anisotropy). Spin-spin coupling, multiplets, spin systems. Protons on heteroatoms. Exchangeable protons. Coupling of protons to other important nuclei. Chemical shift equivalence. Magnetic equivalence (spincoupling equivalence). AMX, ABX, and ABC rigid systems with three coupling constants. Chirality. Vicinal and geminal coupling in rigid systems: Karplus correlations. Long-range coupling. Selective spin decoupling. Double resonance. Nuclear Overhauser Effect (NOE). Shift reagents. Carbon Magnetic Resonance Spectrometry (13C-NMR). Total spin decoupling spectra. Off-resonance spectra. Chemical shift equivalence. Correlation NMR spectrometry. DEPT and APT experiments. Variable temperature NMR. Basics of 1H-NMR and 13C-NMR spectrum interpretation. Mass Spectrometry (MS) Ionization systems: electron impact, chemical ionization, fast atom bombardment (FAB) – ESI – APCi – MALDI. Analyzers: magnetic and electrostatic sectors, quadrupolar systems, ion trap, time of flight systems, orbitrap, tandem mass spectrometry. Molecular ion, exact masses, isotopic abundance, isotopic clusters, elemental composition. Typical fragmentation patterns in mass spectra, McLafferty rearrangement.

Silverstein, Webster, Kiemle. Identificazione spettrometrica di composti organici, Terza Edizione Italiana (2016) Casa Editrice Ambrosiana, Milano.