- Docente: Andrea Zattoni
- Credits: 6
- SSD: CHIM/01
- Language: Italian
- Teaching Mode: Traditional lectures
- Campus: Bologna
-
Corso:
Second cycle degree programme (LM) in
Health Biology (cod. 9212)
Also valid for Second cycle degree programme (LM) in Health Biology (cod. 9212)
Learning outcomes
The student acquires basic knowledge of quantitative analytical methods and common analytical techniques based on biospecific reagents (enzymatic analysis, immunometric methods). The student is able to: solve in the laboratory a clinical analysis problem by choosing one or more methods; critically evaluate the analytical data.
Course contents
INTRODUCTION TO ANALYTICAL CHEMISTRY
The analytical process
Sampling of biological fluids
STATISTICS
Statistical analyisis of replicates
Accuracy and precision
Statistical paramaters
Parametric significance tests: T test, F test, Q test
Confidence interval
Linear regression
Calibration: external standards, internal standard, standard additions
Validation of an analytical method: accuracy, precision (repeatability, reproducibility), sensitivity, limit of detection, limit of quantification, dynamic range, specificity, selectivity, robustness, solidity.
SPECTROPHOTOMETRY
Lambert-Beer law: definition and use in quantitative analysis
Absorbance and transmittance
Absormance measurements: choice of wavelength and deviations from ideality
Instrumentation for spectrophotometry. Components and instrumental setup
CHROMATOGRAPHY
Basics of chromatography: chromatogram, chromatographic peak, retention time, void time
Optimization of chromatographic separations: capacity factor, selectivity, efficiency, resolution
Instrumentation for chromatography
Gas chromatography: instrumentation and methods
Liquid chromatography (HPLC). Instrumentation
Methods in HPLC
Normal phase and reversed phase
Quantitative analysis in chromatography
MASS SPECTROMETRY
Basics and instrumental setup
Hard and soft ion sources
Mass analyzers
MS of proteins
IMMUNOMETRY
Principles and advantages of immunometric methods
Monoclonal and polyclonal antibodies. Properties and applications
Labels for immunometry. Immunoenzymatic methods
Classification of immunomethods
Homogeneous and heterogeneous, competitive and non-competitive methods
Calibration
Quantitative analysis
ENZYMATIC METHODS
Enzymatic analysis. Basics and applications.
Properties and structure of enzymes
Enzymatic activity
Kintics
Regions of the activity/[substrate] curve
Michaelis-Menten equation
Determination of enzymatic activity
Single-point, double-point and multipoint methods.
Quantitative analysis of antigens.
Endpoint methods
Measurement in enzymatic analysis
Readings/Bibliography
L. Spandrio "Principi e Tecniche di Chimica Clinica", Piccin
-A.J. Ninfa, D.P. Ballou "Metodologie di Base per la Biochimica e la Biotecnologia", Zanichelli (consultazione)
-J.M. Berg, J.L. Tymoczko, L. Stryer "Biochimica", V Edizione, Zanichelli
Teaching methods
Class lectures with overhead projection. The slides are available on http://campus.unibo.it
Assessment methods
Final exam on the course program
Teaching tools
Individual laboratory exercises.
Office hours
See the website of Andrea Zattoni
SDGs
This teaching activity contributes to the achievement of the Sustainable Development Goals of the UN 2030 Agenda.