Learning objectives
The course aims to provide students with a thorough knowledge of major
instrumental analytical techniques, including spectroscopic techniques,
separation and hyphenated techniques.
A critical discussion of equipments, procedures and
interpretation of test results in the fiels of biotechnologies is emphasised.
It intends to supply the concepts and abilities that allow the understanding and explanation of the basic principles of analytical chemistry and its importance in biotechnology.
At the end of the course the student is expected to be able to:
- know the fundamental principles of the instrumental analysis methods including the classification of the instrumental analytical techniques and the corresponding selection criteria
- apply the major analytical chemistry techniques
- verbally express the concepts and contents learned
- acquire and use tools to help student learn how to learn, with additional materials for further information
- undertake academic studies of a higher level with a sufficient degree of autonomy.
Prerequisites
General Chemistry; Organic and Bioorganic Chemistry. Statistics
Course unit content
Introduction to instrumental analytical methods.
Classification of analytical techniques.
Criteria for the choice of analytical techniques.
The laboratory process: pre-analytical, analytical and post-analytical phases of testing.
Development and method validation.
Calibration methods.
Spectroscopic techniques.
Fundamentals of spectrophotometry.
Molecular Absorption and Fluorescence Spectroscopy. Infrared Absorption
Spectroscopy.
Atomic Absorption Spectroscopy.
Atomic Emission Spectroscopy.
Mass Spectrometry.
Separation techniques.
Gas chromatography and liquid chromatography.
Full programme
Steps of an analytical process: pre-analytical, analytical and post-analytical phases of testing. 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.
Spectroscopic techniques:
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 and application 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. Quantitative analysis in fluorescence spectroscopy: performance of the technique.
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.
Separation techniques
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.
Qualitative and quantitative analysis in GC and HPLC.
Bibliography
K.A. Rubinson, J.F. Rubinson, Chimica Analitica Strumentale, Zanichelli,
2002.
Holler, Skoog, Crouch, Chimica Analitica Strumentale, II edizione, EdiSES,
2009.
D.S. Hage, J.D. Carr, Chimica Analitica e Analisi Quantitativa, Piccin, 2012.
Teaching methods
Lectures. Classroom exercises designed to develop analytical skills. Interactive teaching.
Blended synchronous learning will be adopted: face-to-face course and in real time, simultaneously transmitting it to students who are remotely accessing the same course (via Teams). Video-recording of each lesson will be available within the week during which it has been delivered.
To promote active participation in the course, the lectures will be accompanied by interactive educational paths. The slides of the Course are considered an integral part of the teaching material.
Assessment methods and criteria
Oral examination in person.
If it will be necessary to integrate the execution of the examinations with the remote modality due to the persistence of the health emergency, we will proceed as follows: oral tests (in person/remotely)
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.
Other information
Illustration of case studies related to the application of instrumental analytical techniques in food, environmental and biological field, with a focus on biotechnology.
