Principle of spectroscopy and applications to quantitative biology
A.A. 2020/2021
Obiettivi formativi
The primary objective of this course is to introduce the basic concepts, principles, and techniques of modern analytical chemistry that would provide students with an analytical mind shape and abilities to solve diverse analytical problems, involved in the biotechnologies, in an efficient, quantitative and qualitative way. One of the aim is to provide to the students adequate preparation in order to identify the different components of the biological system and to convey the importance of accuracy and precision of the analytical results.
Risultati apprendimento attesi
On successful completion of this course, students will be able:
1) to understand the different analytical methods for molecular and supramolecular analysis.
2) to establish an appreciation of the role of chemistry and molecular spectroscopy in quantitative analysis, in measurement and problem solving for analytical tasks.
Moreover students will be able to select molecular probes and to design experiments in order to understand the mechanism of cellular uptake, or to better visualize the different components of biological system.
Through the experimental part of the course, the students will be trained to correctly interpret and to communicate the experimental results.
1) to understand the different analytical methods for molecular and supramolecular analysis.
2) to establish an appreciation of the role of chemistry and molecular spectroscopy in quantitative analysis, in measurement and problem solving for analytical tasks.
Moreover students will be able to select molecular probes and to design experiments in order to understand the mechanism of cellular uptake, or to better visualize the different components of biological system.
Through the experimental part of the course, the students will be trained to correctly interpret and to communicate the experimental results.
Periodo: Primo semestre (se vi sono più edizioni controllare anche il periodo dell'edizione, che può essere diverso)
Modalità di valutazione: Esame
Giudizio di valutazione: voto verbalizzato in trentesimi
Programma e organizzazione didattica
Edizione unica
Responsabile
Periodo
Primo semestre
Programma
The topics are:
- Potentiometric methods: junction potential, reference and indicator electrodes, calibration, direct methods
- Potentiometric titrations
- Infrared spectroscopy: fundamental principles, frequency of infrared absorption and chemical structure.. Instrumentation and recording, IR Spectrophotometer and Fourier Transform IR (FT-IR). Evolution and applications of IR spectroscopy.
- Spectrophotometric methods, the electromagnetic radiation and the light/matter interaction, absorption spectroscopy, chromophores, Lambert-Beer equation
- Spectrophotometric titrations
- Emission spectroscopy: photophysical deactivation paths of the excited-state, kinetics of excited states quenching, excimers and exciplexes. Steady state and time-resolved emission spectroscopy. Fluorescence methods: fluorescent probe for cell imaging. Chemiluminescence
- Nuclear magnetic resonance (NMR): general principles, nuclear properties, magnetic field and electromagnetic radiation (radiofrequency).
Proton nuclear resonance: 1H NMR chemical shift and structure, chemical shift equivalence: random, isochronal protons, homotopic protons, enantiotopic, diasterotopic. Magnetic Equivalence.
Practice on a collection of proton spectra of simple aromatic compounds, spin system simulation (cascade diagrams), reading of spin systems (determination of coupling constants and chemical shift).
NOe differential experiments with examples.
Nuclear magnetic resonance of carbon-13: general principles, relative and absolute sensitivity of an NMR experiment, chemical shifts, proton-proton and proton-carbon coupling, examples of simple aromatic and non-aromatic compounds, 13C-NMR spectra recorded with the APT and DEPT techniques.
Two-dimensional magnetic resonance spectroscopy (2D). COrrelation Spectroscopy COSY, HSQC; HMBC, NOESY and TOCSY.
- Mass Spectrometry: General principles and instrumentation. Sources (EI, CI, FAB, MALDI, ESI APCI), analyzers (magnetic, quadrupole, ion trap), detectors. Analysis of a mass spectrum: molecular ion, fragment ions, general classification of fragmentation reactions. Systematic part: fragmentation reactions of the main classes of organic compounds with exercises.
-Chromatographic techniques
Final Exercises on Structural Determination of Organic Compounds with Combined Techniques.
- Potentiometric methods: junction potential, reference and indicator electrodes, calibration, direct methods
- Potentiometric titrations
- Infrared spectroscopy: fundamental principles, frequency of infrared absorption and chemical structure.. Instrumentation and recording, IR Spectrophotometer and Fourier Transform IR (FT-IR). Evolution and applications of IR spectroscopy.
- Spectrophotometric methods, the electromagnetic radiation and the light/matter interaction, absorption spectroscopy, chromophores, Lambert-Beer equation
- Spectrophotometric titrations
- Emission spectroscopy: photophysical deactivation paths of the excited-state, kinetics of excited states quenching, excimers and exciplexes. Steady state and time-resolved emission spectroscopy. Fluorescence methods: fluorescent probe for cell imaging. Chemiluminescence
- Nuclear magnetic resonance (NMR): general principles, nuclear properties, magnetic field and electromagnetic radiation (radiofrequency).
Proton nuclear resonance: 1H NMR chemical shift and structure, chemical shift equivalence: random, isochronal protons, homotopic protons, enantiotopic, diasterotopic. Magnetic Equivalence.
Practice on a collection of proton spectra of simple aromatic compounds, spin system simulation (cascade diagrams), reading of spin systems (determination of coupling constants and chemical shift).
NOe differential experiments with examples.
Nuclear magnetic resonance of carbon-13: general principles, relative and absolute sensitivity of an NMR experiment, chemical shifts, proton-proton and proton-carbon coupling, examples of simple aromatic and non-aromatic compounds, 13C-NMR spectra recorded with the APT and DEPT techniques.
Two-dimensional magnetic resonance spectroscopy (2D). COrrelation Spectroscopy COSY, HSQC; HMBC, NOESY and TOCSY.
- Mass Spectrometry: General principles and instrumentation. Sources (EI, CI, FAB, MALDI, ESI APCI), analyzers (magnetic, quadrupole, ion trap), detectors. Analysis of a mass spectrum: molecular ion, fragment ions, general classification of fragmentation reactions. Systematic part: fragmentation reactions of the main classes of organic compounds with exercises.
-Chromatographic techniques
Final Exercises on Structural Determination of Organic Compounds with Combined Techniques.
Prerequisiti
Knowledge of the fundamentals of chemistry, organic chemistry, physics and biochemistry.
Metodi didattici
The course is divided into classroom lectures, in which the topics are illustrated both with slides and by carrying out exercises on the blackboard. At the end of the course, an educational experimental laboratory is organized in which the students, organized in small groups, carry out four different experiences concerning both traditional and instrumental quantitative analysis.
Materiale di riferimento
Skoog, West, Holler, Crouch, Fondamenti di Chimica Analitica. EdiSES. (3° ed. italiana 2015)
Slides of the lessons available on Ariel website of the course
Slides of the lessons available on Ariel website of the course
Modalità di verifica dell’apprendimento e criteri di valutazione
The knowledge of the topics reported in the course's program will be evaluated by means of a three hours-written test, which is aimed at verifying the knowledge of the topics. In this test the student is asked to solve three exercises concerning the interpretation of data collected from instrumental analysis of biopharmaceutical molecules and to answer two or three theory questions concerning analytical approaches for the characterization of biomolecules.
Note that unanswered questions will be counted as wrong answers. Examples of problems and questions and their evaluation will be provided during the course.
Note that unanswered questions will be counted as wrong answers. Examples of problems and questions and their evaluation will be provided during the course.
CHIM/01 - CHIMICA ANALITICA - CFU: 0
CHIM/02 - CHIMICA FISICA - CFU: 0
CHIM/03 - CHIMICA GENERALE ED INORGANICA - CFU: 0
CHIM/06 - CHIMICA ORGANICA - CFU: 0
CHIM/02 - CHIMICA FISICA - CFU: 0
CHIM/03 - CHIMICA GENERALE ED INORGANICA - CFU: 0
CHIM/06 - CHIMICA ORGANICA - CFU: 0
Esercitazioni: 16 ore
Lezioni: 72 ore
Lezioni: 72 ore
Docente:
Panigati Monica
Docente/i
Ricevimento:
Tutti i giorni dalle 10,00 alle 17,00
Dipartimento di Chimica - 1° Piano lato A