Master Program in Molecular Biotechnology

Mass Spectrometry

Mass Spectrometry

Academic year 2024/2025

Sommario del corso

• Course objectives
• Results of learning outcomes
• Program
• Course delivery
• Learning assessment methods
• Suggested readings and bibliography
• Teaching tools
• Course card

Course objectives

The teaching is part of the general objective of the course to provide knowledge and skills in the field of analytical biological chemistry, with particular reference to the knowledge and understanding of instrumental analytical methodologies currently in use in the laboratories of chromatography-mass spectrometry:

1) Detailed knowledge of extractive procedures of different classes of biomolecules (based on investigation of structure-chemical physics properties relationships).
2)Learning of instrumental analysis techniques (GC-MS and HPLC-MS).
3)Learning of basic mechanisms of mass spectra interpretation both of electronic impact and electrospray for a reliable identification of biological molecules.
4)Detailed definition of analytical pattern of particular complexity like those concerning the MS analysis of proteins.
Basic knowledge of extractive and separation techniques.
Principles of mass spectrometry. Elements of analytical chemistry,
organic chemistry and biochemistry.

Results of learning outcomes

Knowledge of analysis procedures of different classes of biological molecules. Sample preparation and clean-up, chromatography and MS skills, in particular in impurity profiling and quantitation and in proteomic analysis.

Program

LC-MS concepts. Chemical structure of drugs and biomolecules. ionization of biomolecules, pKa.

Preanalytical phase: Biomolecules concentration level. Extraction, Clean-up. Deproteinization. Matrix effect. Liquid-liquid extraction. Solid Phase Extraction. Immunoaffinity extraction. Molecular imprinted polymers. SPE examples: mixed mode, SCX, SAX, RAM, MIP, low-volume.

Chromatography: principles. Chromatographic parameters: efficiency, selectivity, capacity factor, resolution. Van Deemter equation. GC: derivatization, detectors. LC: instrumentation, stationary and mobile phases, detectors. SFC. CE.

Mass Spectrometry. Introduction. EI fragmentation. Odd and even electron ions. N rule. Sigma bond and alpha cleavage. Rearrangements. Chemical Ionization, PTR, ESI, NSI, APCI, APPI, MALDI, DESI, ambient ionization techniques. ICP-MS. LC-MS fundamentals. LC-MS applications. Peptidomics, Proteomics and Metabolomics.

Course delivery

The course is delivered face-to-face, in two hours lectures. The total course time is 40 hours.

The presentations slides toghether with depth study papers are available in the e-learnig UniTO moodle website.

After the recent emergency situation, the teaching will be made compatible with an integrated learning environment in e-learning mode, suitable for face-to-face situations, those in which teaching is provided entirely online and forms of blended teaching.

Depending on the changing situations, the platform course will welcome: lesson materials (video recordings and / or audio presentations); exercises to be carried out online, but also face-to-face; in-depth teaching materials that can be discussed through a forum and / or in the classroom during the lesson-debate. The objective of the learning environment is to create continuity between the different teaching moments and between the different situations (remote activities and face-to-face activities).

Learning assessment methods

Written test. Multiple questions requiring short and targeted answers (35 questions in 45 minutes).

Depending on the recent emergency situation, the exam may be in oral remote (webex) mode.

Jurgen H. Gross, Spettrometria di massa,  EdiSES 2016 (Mass Spectrometry, Springer-Verlag 2011)

Courses that borrow this teaching

• Mass Spectrometry (BIO0168C Pds 306-TB)
  Master Program in Molecular Biotechnology

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