B030350 - METABOLOMIC TECHNOLOGIES WITH LABORATORY

Italian

Introduction to the Omics
Metabolite Definition
-Gas Chromatography/Mass-Spectrometry (GC/MS); Liquid Chromatography/Mass-Spectrometry (LC/MS); Capillary Electrophoresis (CE-MS)
-Metabolic Profiling Nuclear Magnetic Resonance Spectroscopy (NMR‏; High resolution magic angle spinning (HR-MASS). Collection and handling of different type of samples
Acquisition of the NMR spectra: Bucketing, alignment, normalization, Data scaling
Data Analysis
Application examples in clinical studies

Slides self explaining

Students will acquire knowledge about the Omics and particularly about the metabolomics techniques. They will learn the principles and main techniques for metabolite evaluation in different type of biological sample and the main analytical techniques of metabolomic data analysis. Furthermore, they will learn to apply these techniques to the clinical studies. Students will acquire not only the principles of metabolomic but they apply the techniques during the time for laboratory exercise.

Propedeuticity approved by degree course

Frontal lessons and professional formative activities

For information about the course, please contact the following email:
Prof.Anna Maria Gori annamaria.gori@unifi.it

Prof. Leonardo Tenoritenori@cerm.unifi.it

The oral test will include all the topics of the lessons.

Metabolomic Technologies and Laboratory
Introduction to the Omics: genomics, metagenomics, Epigenomics, Transcriptomics, Proteomics, lipidomics . interactomics etc
Introduction to Metabolomic
Benefits of analyzing the metabolome
Metabolomics is More Time Sensitive Than Other “Omics”
Influence of diet on metabolomics
Metabolite Definition
Metabolomic tecniques application in clinical studies
• Main Analytical Techniques: Gas Chromatography/Mass-Spectrometry (GC/MS); Liquid Chromatography/Mass-Spectrometry (LC/MS); Capillary Electrophoresis (CE-MS)
Chromatography Techniques: column chromatography; Mass Spectrometry (MS)‏
• Metabolic Profiling Methods
• Analytical Techniques:
– Nuclear Magnetic Resonance Spectroscopy (NMR)‏
– High resolution magic angle spinning (HR-MASS)
• Nuclear Magnetic Resonance (NMR): principles
• Magnetic properties of subatomic particles: nuclear spin and nuclides, spin magnetic moment, quantization

Samples for metabolomic: characteristics and handling
 Urine: Urine Collection and Handling
 Serum/plasma: collection in tubes with or without anticoagulants and handling
 Saliva: collection and preparation the same as urine
 Fecal extracts: feces extraction and preparation the same as urine
 Exhaled breath condensate Sample preparation for NMR : the same as urine
 Cells/tissues extracts
 Tears, sweat, vaginal fluid, seminal fluid, synovial fluid, bile, cerebrospinal liquid, .
 Intact tissues with HRMAS
Acquisition of the NMR spectra
From spectrum to data
Data preparation: bucketing, alignment, normalization,
Data scaling

Data Analysis
Univariate statistics
Multivariate statistics:
Unsupervised methods:
- Projection based methods (principal component analysis)
- Cluster analysis (k-means, hierarchical clustering)
Supervised methods
- Projection based methods (LDA, PLS,.)
- Machine learning (SVM, k-nn)

Application examples