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Analytical Chemistry

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Chimica Analitica

Course ID
BIO0133
Teacher
Prof. Claudio Medana
Degree course
[0101M21] Molecular Biotechnology
Year
2nd year
Type
Distinctive
Credits/Recognition
5
Course disciplinary sector (SSD)
CHIM/01 - chimica analitica
Delivery
Formal authority
Language
English
Attendance
Obligatory
Type of examination
Written
Type of learning unit
modulo
Modular course
Chemistry (BIO0133)
Prerequisites
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.
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Sommario del corso

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Learning 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.

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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.

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Syllabus

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.

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Course delivery

5 CFU (40 h) lectures.

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Learning assessment methods

Written examination. Multiple questions requiring short and targeted answers.

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Support activities

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

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Suggested readings and bibliography

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

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Last update: 20/07/2017 15:36
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