00090 - Analytical Chemistry

Learning outcomes

Basic knowledge of conventional spectroscopic and separative analytical techniques and of bioanalytical techniques; ability to interpret analytical data and to chose the most appropriate analytical method.

Course contents

THE ANALYTICAL PROCESS, STATISTICAL TREATMENT OF EXPERIMENTAL DATA, DATA ELABORATION: Errors in quantitative analysis, errors in analytical chemistry, statistics of errors, tests of significance, error propagation, significant figures, absolute and relative analytical methods, calibration (external calibration, standard additions, internal calibration), least squares fitting, characteristics of an analytical procedure (accuracy, precision, specificity and selectivity, linearity range, detection and quantitation limits, robustness, ruggedness).
ANALYTICAL SIGNAL AND INTRODUCTION TO INSTRUMENTAL ANALYSIS
SPECTROSCOPIC METHODS: electromagnetic radiation, molecular absorption spectra in gaseous and condensed state (electronic, vibrational and rotational contributions), absorbance and transmittance, Lambert-Beer law, deviations from the Lambert-Beer law, analysis of mixtures, error in spectrophotometric measurements, spectrophotometers (light sources, filters and monochromators, sample cuvettes, detectors, instrument configurations). FLUORESCENCE AND PHOSPHORESCENCE: emission and excitation spectra, quantitative analysis, atomic spectrometry, atomic absorption and emission spectra, atomizers (flame atomizers, electrothermal atomizer, plasma atomizer), hollow cathode lamp, instrument configurations, spectral and chemical interferences. Polarized and time-resolved fluorescence.
SEPARATIVE TECHNIQUES: methods classification, theory of chromatography, liquid chromatography. Detectors: absorbance (conventional and diode array), fluorescence, refraction index, light scattering, electrochemical. Development and optimization of a chromatographic method.
MASS SPECTROMETRY: soft impact techniques for biomolecules (proteins and nucleic acids) analysis: MALDI-TOF, electrospray. Protome analysis.
ENZYMATIC ANALYSIS: kinetics of enzymatic reactions, Michaelis-Menten equation, enzyme activity measurements: simple and couplet reactions. One point and two points methods. Dose-response relationships, optimization of experimental conditions, treatment of analytical signal, enzyme activity measurements using UV-Vis spectrophotometry, evaluation of metabolite concentrations,  non-spectroscopic techniques.
IMMUNOLOGICAL METHODS: analytical characteristics of reagents for immunoassays (antisera, monoclonal antibodies, immunogens, tracers), use of reagents for immunoassays, thermodynamics of the antigen-antibody reaction, competitive and non-competitive immunoassays, evaluation of the results of competitive and non competitive immunoassays and quantitative analysis, enzyme immunoassays (EIA, ELISA) with spectrophotometric, fluorescent and chemiluminescent detection.
MOLECULAR BIOLOGY TECHNIQUES: kinetics and thermodynamics of nucleic acids interactions, nucleic acids hybridization reactions, direct and indirect labeling of gene probes, detection of hybrids, signal amplification techniques. PCR and real-time PCR. Quantitative analysis.
BIOSENSORS: characteristics of biosensors. Components of a biosensor: types of biospecific elements and signal transducers. Analytical signal. Examples: electrochemical, piezoelecteric and optic biosensors.

Readings/Bibliography

Copy of the slides will be available for the students

Teaching methods

Lectures dealing with the topics of the program.

Assessment methods

Oral exam.

Teaching tools

PC. videoprojector

Office hours

See the website of Aldo Roda