Learning outcomes of the course unit
The course aims to provide students with a thorough knowledge of major
instrumental analytical techniques, including spectroscopic techniques,
separation techniques, hyphenated techniques and electroanalytical
techniques. A critical discussion of equipments, procedures and
interpretation of test results in the fiels of biotechnologies is emphasised.
At the end of the course the student is expected to be able to:
- understand and acquire knowledge on the methods of analytical
chemistry for the study of molecules in their possible applications to
- apply the analytical methodologies and procedures of chemistry
- apply the main analytical chemistry methods
- verbally express the concepts and contents learned
- acquire and use tools to help student learn how to learn, with additional
materials for further information
General Chemistry; Organic and Bioorganic Chemistry. Statistics
Course contents summary
Introduction to instrumental analytical methods.
Classification of analytical techniques.
Criteria for the choice of analytical techniques.
Fundamentals of spectrophotometry.
Molecular Absorption and Fluorescence Spectroscopy. Infrared Absorption
Atomic Absorption Spectroscopy.
Atomic Emission Spectroscopy.
Gas chromatography and
Steps of an analytical process. Introduction to instrumental analytical techniques and their classification. Classification of analytical methods and criteria for choice of methods.
Quality parameters of analytical methods: linearity range of response, limit of detection, limit of quantitation, sensitivity, selectivity, precision, trueness, accuracy.
Methods for determination of concentrations; calibration with external standards.
UV-visible molecular absorption spectroscopy. Relation between the electronic structure and spectral bands. Instrumentation: sources, monochromators, detectors. Photodiode array detector. Definition of transmittance and absorbance. Beer-Lambert law. Applications to quantitative analysis.
Molecular fluorescence spectroscopy. Instrumentation. Effect of temperature on fluorescence. Quantum yield. Examples of fluorescence of biomolecules. Fluorescent probes for protein and DNA.
Infrared absorption spectroscopy. Origin of spectra; relationship between modes of vibration and absorption zones. Instrumentation: FT-IR spectrophotometer, applications to the characterization of organic substances.
Atomic absorption spectrophotometry. Origin of atomic spectra, instrumentation: sources, atomization systems. Nonspectral interferences. Sample treatment for the determination of trace metals. Performance of different atomization systems.
Atomic emission spectrophotometry. Instrumentation: ICP source, high-resolution monochromator, detectors. Performance of the ICP technique for the analysis of elements.
Mass spectrometry. Principles. Instrumentation: sources (electron ionization, chemical ionization, electrospray ionization). Analyzers: quadrupole, ion trap, time of flight. Detectors.
Principles of chromatography. Gas Chromatography and Liquid Chromatography.
Chromatographic parameters. Retention time. Resolution. Efficiency of a chromatographic column.
Gas Chromatography. Mechanisms of interaction in gas-solid and gas-liquid chromatography. Stationary phases. Instrumentation: injectors, columns, detectors. Capillary columns. Universal and selective detectors. Gas chromatography-mass spectrometry. Temperature programming and isothermal operations.
Liquid Chromatography. Mechanisms of separation. Classes of polarity of substances and stationary phases. Adsorption chromatography, partition, ion exchange, molecular exclusion. HPLC instrumentation: columns, pumps, detectors. Isocratic and gradient elution. Liquid chromatography-electrospray-mass spectrometry for biological applications.
K.A. Rubinson, J.F. Rubinson, Chimica Analitica Strumentale, Zanichelli,
Holler, Skoog, Crouch, Chimica Analitica Strumentale, II edizione, EdiSES,
D.S. Hage, J.D. Carr, Chimica Analitica e Analisi Quantitativa, Piccin, 2012.
Academic teaching. Interactive teaching.
Academic teaching will be supported by interactive educational paths
with the aim of stimulating interest in the subject and comparing
different approaches to the topics covered.
The teaching materials used in the classroom are uploaded to Elly at the
beginning of the course. The slides of the Course are considered an
integral part of the teaching material.
e gas-liquido. Fasi stazionarie liquide e solide. Strumentazione: iniettori,
colonne, rivelatori. Colonne capillari. Rivelatori universali e selettivi.
Accoppiamento gascromatografia-spettrometria di massa. Operazioni in
isoterma e in programmata di temperatura.
Cromatografia liquida: meccanismi di separazione su fasi stazionarie
solide e liquide. Classi di polarità di sostanze e di fasi stazionarie.
Cromatografia di adsorbimento, di ripartizione, a scambio ionico, ad
esclusione molecolare. Strumentazione per HPLC, colonne, pompe,
rivelatori. Eluizione in isocratica e in gradiente di eluizione.
Accoppiamento cromatografia liquida-spettrometria di massa per
applicazioni in campo biologico:sorgente electrospray.
Illustration of case studies related to the application of instrumental
analytical techniques in food, environmental and biological field, with a
focus on biotechnology.
Assessment methods and criteria
The oral exam is evaluated on a 0-30 score scale. The final evaluation is communicated immediately at the end of the test itself.
The student is evaluated on the basic concepts of instrumental analytical chemistry, on the acquisition of a formally correct language, and on the development of links between the different parts of the course. It 'also required a critical evaluation of instrumental analytical techniques and proper classification of analytical methods, with critical evaluation of the method quality parameters.
Illustration of case studies related to the application of instrumental analytical techniques in food, environmental and biological field, with a focus on biotechnology.