Analytical Chemistry

FRANCESCA BUIARELLI

The educational objectives that this course intends to achieve are:
- Learning of problems related to sampling and sample preparation techniques prior to the instrumental chemical analysis.
- In-depth knowledge of the main extraction and instrumental separation techniques (chromatographic, electrophoretic, in the supercritical phase and in fractionation in the flow field).
- Knowledge of the parameters necessary for the validation of an analytical method.
At the end of the course, organized in lectures and in parallel to practical laboratory experiences, students will be able to:
- Development, improvement, and validation of an analytical method to perform trace analysis of analytes in different real matrices, planning the extraction, separation and chromatographic detection procedures
- Chromatogram Interpretation
- Performing a qualitative and quantitative analysis with the statistical treatment of the data

Ability to interpret a chromatogram
ability to prepare suitable sample purification
ability to calculate validation parameters
ability to choose the appropriate instrumental method of analysis

The course "INSTRUMENTAL ANALYTICAL CHEMISTRY II WITH LABORATORY" is included in the master's course, at the first year of "Analytical Chemistry". The necessary knowledge, acquired in the three-year degree, are provided by the basic Chemistry, Analytical Chemistry, Organic Chemistry and Chemistry-Physics teachings.

Principles and theory of the separation science and chromatography. Theory of extraction and the separation science. Nernst's law. Partition coefficient. Extraction of acids and bases. Distribution ratio. Extraction of charged molecules. Extractors for solid and liquid. Application in the environmental field. Chromatographic pararameters: Net and specific retention volume. Capacity ratio. Separation factor. Theoretical plates. Van Deemter's equation. Resolution. Theory and principles of thin layer chromatography. Gas chromatography: gas-solid. Stationary phases. Gas-liquid chromatography: supports, stationary phases and their classification. Scheme and components of a gas chromatograph: carrier gas, Injectors, packed columns and capillary columns, chemically linked phases. Van Deemter's equation for packed columns and capillaries. Conductivity detectors, flame ionization, electron capture, flame photometry and thermionic emission. Compressibility factor, retention indices. Introduction to high-resolution chromatography (HPLC). Adsorption chromatography. Adsorbents. Adsorption isotherms. Eluotropic series of solvents. Properties of solvents, viscosity and solubility of gases in solvents, filtration of solvents and their degassing. Liquid-liquid chromatography. Supported phases. Reversed phases: preparation, properties, end capping, choice of the mobile phase. Modified phases: cyan, amine. Ionic exchange: classification, synthesis, swelling, exchange capacity, selectivity. Size exclusion chromatography: permeation mechanism, gel classification, choice of mobile phase, molecular weight determination. Chiral and affinity chromatography. Modern stationary phases and new trends in HPLC. Isocratic, step, gradient elution. HPLC of small molecules and biological macromolecules. Scheme and components of a high-resolution liquid chromatograph: solvents, pumps, mixers, injectors, columns and their classification with respect to the internal diameter. Refractive index detectors, spectrophotometric, spectrofluorimetric, light scattering, overview of electrochemical and conductivity detector.
Troubleshooting in GC and HPLC
Capillary electrophoresis. Principles, parameters influencing resolution. Electrophoretic techniques and their classification: gel, capillary, electrochromatography. Instrumentation.
Troubleshooting in GC and HPLC Coupling of separative techniques. Two-dimensional chromatography
The principles of "green analytical chemistry".
Development and validation of a method. Accreditation of a laboratory.
Nod of GC-MS coupling, HPLC-MS, electrophoresis-mass spectrometry.
Field flow fractionaction and supercritical fluid chromatography

Qualitative / quantitative analysis and applications Quantitative analysis: calibration line, direct calibration, internal standardization, addition of the internal standard, method of standard additions.
Sample preparation: Purification schemes of real matrices. Strategies for the analysis and purification of organic molecules from complex matrices. Preparative chromatography.

Applications: separation of proteins, peptides, amino acids, studies of metabolism and biotransformation, metabolic and metabolomic profiles. Check purity and product structure. Determination of contaminants in food and in environmental matrices

Lesson slides are available
Practical Gas Chromatography A comprehensive reference Katja Dettmer-Wilde, Werner Engevald Editors. Springer 2014
Essential in Modern HPLC separation Serban C Moldoveanu Victor David Elsevier 2 edition 2022
HPLC and UHPLC for practicing scientist Michael W Dong Wiley Sample preparation techniques in Analytical Chemistry Somenath Mitra Wiley Interscience 2019
Chromatography (Basic Principles, sample preparation and related methods). Elsa Lundanes, Leon Reubsaet, Tyge Greibrokk. Wiley 2013

GAS CROMATOGRAFIA- UNA INTRODUZIONE- Luigi Mondello
http://www.sepsci.unime.it/wp-content/uploads/2015/07/Dispensa-cromatografia.pdf
Modern HPLC for scientist MW Dong Wiley
https://onlinelibrary.wiley.com/doi/pdf/10.1002/0471973106.fmatter

The course includes 56 hours of frontal teachin and 24 hours of laboratory

Frontal lessons
Section (10 hours): Principles and theory of separation science and chromatography

Section II (30 hours): Separative techniques

Section III (10 hours):Instrumentations and main troubleshooting

Section IV (6 hours) Qualitative / quantitative analysis and applications
Sample preparation: Purification schemes of real matrices. Strategies for the analysis and purification of organic molecules from complex matrices

Laboratory exercises and practice in HPLC-UV and GC-FID and sample preparation (24h)

Attendance is useful, not mandatory, but free. Students are invited to participate and take an active part in the lessons.
In order to make the content more comprehensible, slides are projected.
Attendance at least to didactic experiences is strongly recommended

The exam test aims to verify the level of knowledge and in-depth examination of the topics of the teaching program and the reasoning skills developed by the student. The evaluation is expressed in thirtieths (minimum grade 18/30, maximum mark 30/30 with honors).

The evaluation consists of an oral exam (three questions on three different parts of the program). The exam allows you to verify the achievement of the objectives in terms of knowledge and skills acquired as well as communication skills. Very important is the property of language, the clarity of exposition and the critical capacity shown for the resolution of real analytical problems

oral open questions on the different parts of the program, but with demonstration on paper.
Exercises on chromatographic parameters and solving real problems