Evaluation of _1hn1H-NMR and GC-MS-based metabonomics for the assessment of liver and kidney toxicity [Elektronische Ressource] / vorgelegt von Maximilian Sieber

Max Brand

Sujets

• Biologie

Informations

• Publié par: julius-maximilians-universitat_wurzburg
• Publié le: 01 janvier 2009
• Nombre de lectures: 8
• Langue: English
• Poids de l'ouvrage: 6 Mo

Extrait

1Evaluation of H NMR- and GC/MS-based metabonomics for the assessment
of liver and kidney toxicity
Dissertation zur Erlangung des
naturwissenschaftlichen Doktorgrades
der Julius-Maximilians-Universität Würzburg
vorgelegt von
Maximilian Sieber
aus Nürnberg
Würzburg, 2009Eingereicht am: .......................................
bei der Fakultät für Chemie und Pharmazie
1. Gutachter:...................................................
2. Gu......
der Dissertation
1. Prüfer:.........................................................
2. Pr.......................
3. Pr......................................
des öffentlichen Promotionskolloquiums
Tag des öffentlichen Promotionskolloquiums: .....................................................
Doktorurkunde ausgehändigt am:........................................Table of contents
1 Introduction.........................................................................................................1
2 Background.........................................................................................................2
2.1 Conventional methods for the non-invasive detection of nephro- and hepatotoxicity2
2.1.1 Clinical chemistry parameters..............................................................................2
2.1.2 Histopathology.....................................................................................................2
2.1.3 Novel protein biomarkers for kidney and liver injury.........................................3
2.2 Definitions and terminology........................................................................................4
2.2.1 Omics and metabolites.........................................................................................4
2.2.2 Metabonomics vs. metabolomics.........................................................................5
2.3 Analytical platforms....................................................................................................6
12.3.1 H NMR for metabonomic analysis.....................................................................6
2.3.2 GC/MS analysis for metabonomic analysis.........................................................7
2.3.3 LC/MS analysis for metaic analysis.........................................................9
2.3.4 Combination of analytical platforms...................................................................9
2.4 Multivariate data analysis..........................................................................................10
2.4.1 Unsupervised models: principal component analysis........................................10
2.4.2 Supervised models: orthogonal projection to latent structures..........................12
2.4.3 Binning, peak picking and alignment................................................................13
2.4.4 Normalization and scaling.................................................................................14
2.5 Applications in toxicology.........................................................................................15
12.5.1 H NMR-based metabonomics in toxicology....................................................15
2.5.2 GC/MS-based meics in toxicology......................................................15
2.5.3 LC/MS-based metabonomics in toxicology.......................................................16
2.5.4 Identification of putative biomarkers.................................................................16
2.5.5 Confounders.......................................................................................................16
2.5.6 Multiple compound studies................................................................................16
2.6 Model compounds for nephro- and hepatotoxicity....................................................17
2.6.1 Gentamicin.........................................................................................................17
2.6.2 Ochratoxin A......................................................................................................18
2.6.3 Aristolochic acid..............................19
2.6.4 Furan..................................................................................................................21
2.6.5 The InnoMed PredTox project...........................................................................22
3 Objectives..........................................................................................................24
4 Materials and methods.....................................................................................26
4.1 Chemicals and solvents.............................................................................................26
4.2 Animal handling........................................................................................................26
4.3 Clinical chemistry......................................................................................................26
4.4 Study design..............................................................................................................27
4.4.1 Gentamicin...................27
4.4.2 Ochratoxin A......................................................................................................28
4.4.3 Aristolochic Acid...............................................................................................28
4.4.4 Furan..................................................................................................................29
4.4.5 InnoMed PredTox..............................................................................................29
4.5 GC/MS analysis.........................................................................................................314.5.1 Sample treatment for GC/MS analysis..............................................................31
4.5.2 GC/MS analysis.................................................................................................31
4.5.3 Raw data handling and statistical analysis.........................................................32
14.6 H NMR analysis.......................................................................................................33
14.6.1 Sample treatment for H NMR analysis.............................................................33
14.6.2 H NMR analysis...............................................................................................33
4.6.3 Raw data handling and statistical analysis..............34
4.7 LC/MS analysis.........................................................................................................34
4.7.1 Sample treatment for LC/MS analysis...............................................................34
4.7.2 LC/MS analysis..................................................................................................35
4.7.3 LC/MS raw data handling and statistical analysis.............................................35
4.7.4 Targeted bile acid screening..............................................................................36
5 Method and workflow development................................................................37
6 Gentamicin........................................................................................................39
6.1 Introduction...............................................................................................................39
6.2 Results.......................................................................................................................41
6.3 Discussion..................................................................................................................46
7 Ochratoxin A.....................................................................................................50
7.1 Introduction50
7.2 Results.................................................52
7.3 Discussion..........58
8 Aristolochic Acid..............................................................................................66
8.1 Introduction...............................................................................................................66
8.2 Results.......................................................................................................................68
8.3 Discussion..................................................................................................................70
9 Furan..................................................................................................................74
9.1 Introduction74
9.2 Results and discussion...............................................................................................74
10 Innomed PredTox...........................................................................................77
110.1 Single study analysis with H NMR........................................................................78
10.1.1 FP004BA..........................................................................................................78
10.1.2 FP005ME.........................................................................................................81
10.1.3 FP007SE..........................................................................................................82
10.2 Single Study analysis with GC/MS.........................................................................86
10.2.1 FP004BA..............................................................................