ANALYTICAL TECHNIQUES FOR BIOMOLECULES - MOD.2

The Analytical Techniques for Biomolecules course is part of the core educational activities of the Biomolecular Chemistry track within the Master's Degree Programme in Chemistry and Sustainable Technologies. The course is structured into two modules (referred to as Module 1 and Module 2), each comprising theoretical lectures (held in the classroom) and laboratory sessions. Both modules share the overarching goal of building broad knowledge and skills on the principles and applications of the most significant advanced techniques in the bioanalytical field, specifically:
- understanding the principles underlying bioanalytical techniques;
- developing the ability to propose, design, and manage new approaches in the bioanalytical field to solve complex problems related to the qualitative and quantitative determination of chemical species of interest in real matrices;
- acquiring skills to extract and communicate analytical information from experimental data, with full awareness of the capabilities, limitations, and uncertainties associated with techniques, application methods, analytes, matrices, and specific objectives;
- developing flexible and adaptive skills in the use of software tools for managing advanced instrumentation and processing chemical-analytical data, with a focus on automation, big data, and interoperability.

The expected learning outcomes are defined according to the specific content of the two modules that make up the Analytical Techniques for Biomolecules course.

The course aims to provide the foundational knowledge and methodological skills necessary for the development and application of advanced analytical procedures for the qualitative and quantitative determination of biomolecules in complex matrices. Students will acquire a broad range of knowledge and competences in the bioanalytical field, including:

Knowledge and Understanding
- Understanding the fundamental principles and practical adaptations of experimental planning strategies in advanced bioanalytical contexts.
- Understanding the fundamental principles and terminology of advanced analytical techniques.
- Understanding, planning, and applying sample preparation methodologies aimed at the determination of biomolecules in complex matrices.
- Understanding the operational principles (hardware) and software management of advanced analytical instrumentation.

Applying Knowledge and Understanding
- (Understanding the fundamental principles and practical adaptations of experimental planning strategies in advanced bioanalytical contexts).
Ability to define objectives, needs, and specific limitations of advanced experimental activities in the bioanalytical field. Ability to identify, structure, and optimise experimental procedures using advanced design tools.
- (Understanding the fundamental principles and terminology of advanced analytical techniques).
Ability to correctly understand and use the technical-scientific terminology of advanced bioanalytical methodologies. Ability to identify the most suitable technique and methodology for the qualitative/quantitative determination of interest. Ability to define the ideal combination of advanced bioanalytical techniques to obtain comprehensive and integrated information depending on the objective.
- (Understanding, planning, and applying sample preparation methodologies aimed at the determination of biomolecules in complex matrices).
Ability to identify and plan the most appropriate pre-analytical procedures for biomolecule determination based on analytical objectives and constraints, with particular attention to analyte preservation, specificity/selectivity/sensitivity of the analytical technique employed, and contamination.
- (Understanding the operational principles (hardware) and software management of advanced analytical instrumentation).
Ability to operate advanced bioanalytical instrumentation with adaptability to varying and novel configurations.

Judgement Skills
Full awareness of the quality and interpretative value of analytical data obtained or obtainable in response to specific and complex applied problems or questions.

Communication Skills
Ability to communicate effectively, both orally and in writing, with audiences of all levels (general public, non-specialist stakeholders and decision-makers, technicians, experts) using appropriate language to explain the experimental design, the functioning and settings of instrumentation, and the results obtained through the application of advanced analytical techniques and methods for the determination of biomolecules in complex matrices.

Learning Skills
Ability to apply the scientific method as a working tool, both for continuing education at the third cycle level and for professional practice. Ability to identify and apply suitable advanced analytical methodologies to solve the proposed questions.

Basic knowledge of analytical chemistry and statistics.

Omics sciences. Principles of experimental design applied to bioanalytical sciences. Validation of bioanalytical methods, data processing, and data quality. Principles of multivariate analysis. Sampling activities and pre-analytical methods for sample preparation aimed at the determination of biomolecules. Gas chromatography, liquid chromatography, and advanced pseudo-chromatographic techniques applied to bioanalysis. Principles of mass spectrometry. Ionisation sources: EI, ESI, APCI, APPI, MALDI, ICP. Low- and high-resolution mass analysers: time-of-flight; quadrupoles; quadrupole, linear, and orbital traps; magnetic, electrostatic, and double-focusing sectors; hybrid systems. Data acquisition modes in mass spectrometry (full-scan, SIM), interpretation of mass spectra, and applications in inorganic and organic mass spectrometry. Coupled mass spectrometry techniques: gas chromatography and liquid chromatography (IC, LC, (U)HPLC); multidimensional chromatography. Analytical strategies and applications of mass spectrometry for biomolecules: targeted, untargeted, proteomics, sequencing, metabolomics. Principles of bioimaging. Inorganic mass spectrometry (ICP) for metallomics applications: coupling with HPLC and bio-elemental imaging via laser ablation. Applications in biomedical/pharmaceutical analysis, bio-nanomaterials, and the agri-food sector. Theoretical case studies (presentation and discussion of scientific publications) and experimental case studies (laboratory activities).

Lecture notes, slides, scientific publications as case studies provided by the teacher.

The exam consists in an oral discussion. The exam consists of an initial section in which the student is asked to briefly and critically present a literature case study of their choice, relevant to the topics covered in the course. This is followed by in-depth questions on the presented case study and then on other topics discussed during the course. During the oral exam, the following will be assessed:
- theoretical knowledge on advanced bio-analytical techniques and methodologies, both instrumental and pre-analytical.
- technical knowledge on the usage of advanced analytical instrumentation.
- technical knowledge in analytical problem solving.
- theoretical-technical knowledge in the treatment and evaluation of analytical data.

The final mark of the Analytical Techniques for Biomolecules course is single, corresponding to the arithmetic average of the individual marks achieved in the two modules.

oral

Assessment Grid:
27–30: Full mastery of the topics covered in the course; ability to independently reorganise acquired knowledge to adapt the analytical strategies presented in the programme to specific applications; appropriate use of technical terminology.
23–26: Good understanding of the topics covered; fair ability to organise information and effectively present analytical strategies previously described during the course; familiarity with technical terminology.
18–22: Superficial knowledge of the topics covered; partially structured oral presentation requiring occasional guidance; inconsistent use of technical terminology.
Honours (cum laude): awarded in cases of demonstrated ability to rework and expand upon the analytical solutions presented in the programme in a personal, original, and critical manner.

Classroom lessons and laboratory activity.

This subject deals with topics related to the macro-area "Human capital, health, education" and contributes to the achievement of one or more goals of U. N. Agenda for Sustainable Development