66610 - Bioanalytical Chemistry

Learning outcomes

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

Course contents

Room lectures (4 CFU, 32 hours)

- Treatment of quantitative analytical data (6 hours). Errors in experimental measurements; applications of statistics to the representation and interpretation of experimental data; tests of significance; significant digits; propagation of errors.

- Quantitative analysis, calibration, and validation (6 hours). Construction of a calibration curve; linear, linearized, and non-linear calibration curves; principles of linear and non-linear regression; evaluation of the quality of a calibration curve; calibration methods: external calibration, internal calibration, calibration with standard addition; validation of an analytical method; characteristics of an analytical method (accuracy, precision, selectivity, linear and dynamic interval, 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: electrochemical processes, Nernst equation, 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, liquid chromatography (HPLC), gas-chromatography (GC); 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.

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 lectures, supplementary material, instructions for laboratory exercises and drafts of laboratory reports will be available on “Virtuale” (https://virtuale.unibo.it, access with credentials reserved for students enrolled at the University of Bologna).

Suggested textbooks
- J.F. Holler, S.R. Crouch. Fondamenti di chimica analitica, 3° Ed. (EdiSES, 2015).
- D.C. Harris. Chimica analitica quantitativa, 3° Ed. (Zanichelli, 2017).
- 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).
- 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 room lectures and laboratory exercises. During the lectures the topics of the course will be presented and discussed, with theoretical insights and explanatory examples. The laboratory exercises 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.

Due to the type of activity, the attendance of the laboratory requires the performance by the student of the specific training on safety and health in study places (modules 1 – 3). Modules 1 and 2 are in e-learning mode, while information on dates and methods of attendance of module 3 can be found in the specific section of the degree program website.

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 reports is mandatory to be admitted to the final exam.

Teaching tools

Video projector for room 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).

Office hours

See the website of Massimo Guardigli