Academic year 2021/2022
- Course ID
- Prof. Claudio Medana
- Modular course
- Chemistry (BIO0133)
- Degree course
- [0101M21] Molecular Biotechnology
- 2nd year
- Course disciplinary sector (SSD)
- CHIM/01 - chimica analitica
- Formal authority
- Type of examination
- Basic knowledge of extractive and separation techniques.
Principles of mass spectrometry. Elements of analytical chemistry,
organic chemistry and biochemistry.
- Propedeutic for
- This teaching is a point of arrival. It is not propaedeutical.
Sommario del corso
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.
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.
5 CFU (40 h) lectures.
Learning assessment methods
Written examination. Multiple questions requiring short and targeted answers.
The presentations slides toghether with depth study papers are available in the e-learnig UniTO moodle website.
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.
Suggested readings and bibliography
Jurgen H. Gross, Spettrometria di massa, EdiSES 2016 (Mass Spectrometry, Springer-Verlag 2011)