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 is emphasised.
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 methods.
Calibration methods: external standard method, standard addition
method, internal standard method.
Fundamentals of spectrophotometry.
Molecular Absorption and Fluorescence Spectroscopy. Infrared Absorption Spectroscopy.
Atomic Absorption Spectroscopy.
Atomic Emission Spectroscopy.
Gas chromatography and
Steps of an analytical process. Sampling. 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, the standard method of addition; use of internal standards.
UV-visible molecular absorption spectroscopy
Relation between the electronic structure and spectral bands. Instrumentation: sources, monochromators, detectors. Array detector diodes. 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. Non-spectral interferences. Sample treatment for the determination of trace metals. Performance of different atomization systems.
Atomic emission spectrophotometry
Instrumentation: ICP source, monochromator, high-resolution detectors. Performance of the technique for the analysis of elements.
Principles. Instrumentation: sources (electron ionization, chemical ionization, electrospray ionization). Analyzers, magnetic sector, quadrupole, ion trap, time of flight. Detectors.
Principles of chromatography. Gas Chromatography and Liquid Chromatography.
Chromatographic parameters. Retention time. Corrected retention time. Capacity factor. Separation factor. Resolution. Efficiency of a chromatographic column.
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. Internal standard method for quantitative determinations.
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,
Academic teaching. Interactive teaching
Assessment methods and criteria
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.
Slides of the course available to students.