23737 - CHIMICA BIOANALITICA

Learning outcomes

At the end of the course the student will know the basics of conventional spectroscopic and separative analytical techniques, as well as of the main bioanalytical methods. He will be able to select the most appropriate (bio)analytical method to solve a given analytical question, to apply (bio)analytical methods following the appropriate analytical protocols, and to analyze and interpret their analytical results.

Course contents

The course is divided into two teaching modules, in which the topics listed below will be addressed.

Module 1 - Lectures (4 CFU, 32 hours)

Applications of statistics to the processing and interpretation of experimental data (6 hours). Error in experimental measurements; random, systematic, and gross errors; characteristics and identification of different types of errors; statistics on small data sets; confidence interval; statistical significance tests; significant figures; error propagation; rounding.

Quantitative analysis, calibration, and validation (6 hours). Construction of a calibration curve; linear, linearized and nonlinear calibration curves; linear and nonlinear regression analysis; evaluation of the quality of a calibration curve; calibration methods: external, internal and standard-added calibration; validation of an analytical method; characteristics of an analytical method: accuracy, precision, selectivity, linear range and dynamic range, LOD, LOQ, robustness.

Instrumental analytical methods (10 hours). UV-Vis spectrophotometry: electromagnetic spectrum and radiation/matter interaction, light absorption, Beer-Lambert law and limits of validity, spectrophotometric error, quantitative analysis, analysis of mixtures, instrumentation for absorption measurements; fluorimetry: light emission, fluorescence and phosphorescence, excitation and emission spectra, quantitative analysis, instrumentation for emission measurements; potentiometric methods: classical and ion-selective electrodes, glass pH electrode; chromatographic methods: classification of chromatographic methods, theoretical aspects of chromatography, development and optimization of a chromatographic method, planar (es. TLC) and column liquid-phase chromatography, high-performance liquid chromatography (HPLC), HPLC instrumentation; basics of mass spectrometric techniques: mass and fragmentation spectra, identification of analytes, instrumentation.

Bioanalytical methods (10 hours). Enzymatic methods: enzyme kinetics, Michaelis-Menten equation, simple and coupled enzymatic reactions, kinetic and end-point methods, measurement of enzymes, enzyme substrates and other species, detection of products (direct spectrophotometric methods, methods based on chemically- or enzymatically-generated derivatives, fluorometric methods, non-spectroscopic methods); immunological methods: antigen-antibody reactions, labeled and unlabeled immunological methods, competitive and noncompetitive immunological methods, homogeneous and heterogeneous immunological methods, production and characterization of antibodies, tracers, immobilization techniques, detection techniques (colorimetry, fluorescence and chemiluminescence), development and optimization of an immunological method.

Module 2 - Laboratory exercises (2 CFU, 30 hours)

Laboratory exercises with application of analytical (spectrophotometric, potentiometric, and separative) and bioanalytical (enzymatic and immunological) techniques treated in the theoretical lessons. The laboratory exercises include the analysis of an unknown sample and the elaboration of the experimental results to obtain its concentration.

Readings/Bibliography

PowerPoint slides of the lectures, supplementary teaching material (e.g., insights, exercises, multiple-choice auto valuation tests), instructions for laboratory exercises and for writing reports will be available on "Virtuale" (https://virtuale.unibo.it, access with credentials reserved for students enrolled at the University of Bologna).

Recommended reference textbooks

Statistics and processing of experimental data: G. Filatrella, P. Romano. Elaborazione statistica dei dati sperimentali, 2° Ed. (EdiSES, 2022); M. Grotti, F. Ardini. Il laboratorio di chimica analitica, 1° Ed. (EdiSES, 2022).

General analytical chemistry: J.F. Holler, S.R. Crouch. Fondamenti di chimica analitica, 3° Ed. (EdiSES, 2015); D.C. Harris. Chimica analitica quantitativa, 3° Ed. (Zanichelli, 2017).

Bioanalytical chemistry: A. Manz, P.S. Dittrich, N. Pamme, D. Iossifidis. Bioanalytical chemistry, 2° Ed. (Imperial College Press, 2015).

Teaching methods

The course is made up of two modules (lectures and laboratory exercises). During the lectures module the topics of the course will be presented and discussed, with theoretical insights and explanatory examples. The laboratory exercises module will allow each student to acquire the knowledge of instrumental (bio)analytical techniques and the necessary skills for working in a laboratory according to quality and safety guidelines. Laboratory exercises include the analysis of unknown samples: the student will process the results using the tools learned in the course and will submit a report for each experience.

Attendance at laboratory exercises is mandatory (it is necessary to participate in at least 80% of the laboratory activities).

As concerns the teaching methods of this course unit, all students (including all the international incoming exchange students, i.e. ERASMUS) must attend Modules 1 and 2 online (https://www.unibo.it/it/servizi-e-opportunita/salute-e-assistenza/salute-e-sicurezza/sicurezza-e-salute-nei-luoghi-di-studio-e-tirocinio), while Module 3 on health and safety is to be attended in class. Information about Module 3 attendance schedule is available on the website of your degree programme ("studiare"--"formazione obbligatoria su sicurezza e salute").

Assessment methods

Verification of knowledge on the whole program by means of the following tests.

Oral exam on the topics covered during the course, aimed at verifying the acquisition of knowledge and expected skills. In particular, the knowledge of procedures for processing and evaluating experimental data and the understanding of the principles and procedures of the analytical and bioanalytical instrumental techniques will be evaluated.

Evaluation of the laboratory exercises conducted based on the reports submitted by the student. Both the correctness of the procedures used for the elaboration of the experimental data and the concordance of the results with the concentrations of the unknown samples will be considered.

The final grade will be the sum of the marks obtained in the oral exam (maximum 20/30) and in the laboratory exercises (maximum 10/30). The submission of laboratory reports according to the methods that will be illustrated during the laboratory exercises is essential for taking the final exam.

Students with learning disorders and\or temporary or permanent disabilities must contact the office responsible (https://site.unibo.it/studenti-con-disabilita-e-dsa/en/for-students) as soon as possible so that they can propose acceptable adjustments. The request for adaptation must be submitted in advance (15 days before the exam date) to the lecturer, who will assess the appropriateness of the adjustments, considering the teaching objectives.

Teaching tools

Video projector for lectures and analytical laboratory equipped with the necessary instrumentation. The teaching material will be available on “Virtuale” (https://virtuale.unibo.it, access with credentials reserved for students enrolled at the University of Bologna). The instruction for laboratory exercises must be printed and brought to the laboratory.

Office hours

See the website of Massimo Guardigli