Ateneo - UPO

The course will provide the foundations of the most commonly used instrumental analytical techniques in the clinical laboratory (liquid chromatography, gas chromatography, electrophoretic techniques, elemental analysis, etc.), along with the most commonly used sample pretreatment and analysis methods in the clinical laboratory. Performance indicators for analytical methods (LOD, LOQ, recovery, dynamic and linear range, robustness, measurement uncertainty, accuracy, repeatability, etc.) and diagnostic methods (sensitivity, specificity, accuracy, ROC curve) will also be covered. Two credits will be dedicated to laboratory work with small-group experiences using advanced instrumentation: UV-visible spectroscopy and liquid chromatography coupled to high-resolution mass spectrometry.

Material from the teacher

The course aims to provide students with knowledge of the most common analytical instruments (liquid chromatography, gas chromatography, electrophoretic techniques, elemental analysis) and the parameters for evaluating the performance of an analytical method. Knowledge of sample pretreatment methods for the clinical laboratory and the fundamentals of analytical methods used for the determination of blood and urinary parameters are also provided. Finally, knowledge of the performance of diagnostic methods (sensitivity, specificity, ROC curve, accuracy) will be provided. Students will develop critical thinking skills, refining the ability to choose the best experimental strategy to solve a given analytical problem. Communication skills will also be developed through the use of appropriate vocabulary. Two credits will be dedicated to laboratory experiences in small groups during which students will learn basic analytical laboratory operations and conduct experiments using UV-visible spectroscopy and liquid chromatography coupled to mass spectrometry.

None

Lectures, classroom discussions with students, experiences in lab in small groups

Learning will be evaluated through classroom and lab discussions with the students. Students with physical disabilities, Learning Disabilities or Special Education Needs can request specific services and tools via the Staff Sviluppo e Coordinamento Carriere e Servizi alle Studentesse e agli Studenti, consulting the University webpage: https://www.uniupo.it/en/services/servicesstudents-physical-or-learning-disabilities. Students with disabilities, learning disabilities or special education needs, once they have contacted the University Staff, can refer to the tutor in charge of the course to define the examination modalities, concerning academic aspects.

Oral exam consisting of three open-ended questions: one on the theoretical and practical aspects of the analytical techniques covered in class; one on the determination of analytical parameters in biological samples; and one on figures of merit in analytical chemistry. A group report on the experiments performed during the laboratory portion will also be evaluated.
The laboratory report may impact the exam grade by a maximum of 2 points.
The exam is considered passed by demonstrating: knowledge of the general principles of the instrumental techniques illustrated; knowledge of the general principles of determining routine clinical parameters; ability to illustrate at least a basic level of the figures of merit discussed in class; and ability to write a technical report on the experiments performed in the laboratory.
Excellence can be achieved by demonstrating critical thinking during oral questions, specifically: ability to compare different techniques; ability to discuss the details of determining clinical parameters of biological samples; demonstrating in-depth knowledge of how to calculate and evaluate figures of merit in analytical chemistry; and writing a detailed report on the laboratory activities and demonstrating critical thinking.

Part I – Instrumental Analysis Techniques. Introduction to spectroscopic techniques, electromagnetic radiation, absorption, emission, fluorescence, phosphorescence. Molecular UV-visible absorption: principles, instrumentation, and applications. Elementary techniques: introduction, flame and furnace atomic absorption, instrumentation, interference, correction methods, and applications; inductively coupled plasma, instrumentation, and applications. Chromatographic techniques: introduction, plate and velocity theory, liquid chromatography (mechanisms, instrumentation, and applications), gas chromatography (mechanisms, instrumentation, and applications). Electrophoretic techniques: capillary electrophoresis, electrophoretic and non-electrophoretic immunoprecipitation techniques, gel techniques, and applications. Performance indices of an analytical method (LOD, LOQ, recovery, dynamic and linear range, robustness, measurement uncertainty, accuracy, repeatability, etc.). Part II – Analysis of Biological Samples. Sample collection and storage methods. Analytical determinations in the clinical chemistry laboratory: electrolyte balance, trace elements, calcium, magnesium, and major cations, lipids and lipoproteins, protein metabolism, bilirubin and jaundice-related molecules, carbohydrate metabolism, bone remodeling, enzymes, circulating markers, hormones, urinalysis. Screening and confirmatory tests: sensitivity and specificity, accuracy, ROC curve. Part III – Small-group laboratory experiments. Determination of protein content using the Bradford assay. Extraction of lipids and small molecules from biological matrices. Determination of analytes in biological matrices using liquid chromatography coupled to mass spectrometry. Determination of elements in biological samples using ICP-MS and ICP-OES.

Knowledge and understanding - knowledge of the theoretical and theoretical/practical bases of the most modern analysis techniques (chromatography, elemental methods, UV-visible spectroscopy, electrophoretic techniques, basics of mass spectrometry) - knowledge of pretreatment and analysis methods of biological samples - knowledge of figures of merit in analytical chemistry to compare the performance of analytical methods Ability to apply knowledge and understanding - knowing how to compare different analytical techniques - knowing how to choose the best method for determining analytical parameters in biological samples - knowing how to comment on a ROC curve and compare screening methods and confirmatory methods - knowing how to perform the basic operations in the analytical chemistry laboratory - knowing how to carry out simple laboratory experiments with analytical instrumentation Communication skills - acquiring and knowing how to use appropriate vocabulary in relation to the topics covered in the course - knowing how to report the results of an experiment in the form of a written report Autonomy of judgment - ability to choose between different analytical techniques for the same purpose