Biotransformation and toxicokinetics of acrylamide in humans [Elektronische Ressource] / vorgelegt von Eva Katharina Kopp

Lgl-Kopp1

Sujets

• Gesundheit

Informations

• Publié par: julius-maximilians-universitat_wurzburg
• Publié le: 01 janvier 2009
• Nombre de lectures: 27
• Langue: English
• Poids de l'ouvrage: 3 Mo

Extrait

Biotransformation and Toxicokinetics
of Acrylamide in Humans





Dissertation zur Erlangung des
naturwissenschaftlichen Doktorgrades
der Bayerischen Julius-Maximilians-Universität Würzburg





vorgelegt von
Eva Katharina Kopp
aus Starnberg



Würzburg 2009








Eingereicht am:..........................................
bei der Fakultät für Chemie und Pharmazie



1. Gutachter: ..................................................
2. Gutachter: ..................................................
der Dissertation


1. Prüfer: .........................................................
2. Prüfer: .........................................................
3. Prüfer: .........................................................
des Öffentlichen Promotionskolloquiums


Tag des Öffentlichen Promotionskolloquiums: ..........................

Doktorurkunde ausgehändigt am: .....................



Table of Contents I
TABLE OF CONTENTS

Table of Contents ................................................................................................................... I
Abbreviations ....................................................................................................................... V
1 Introduction ...................................................................................................................... 1
2 Background 5
2.1 Physical and Chemical Characteristics of Acrylamide ................................. 5
2.2 Biotransformation and Disposition of Acrylamide ........................................ 6
2.2.1 Biotransformation in Rodents ...................................................................... 6
2.2.2 Biotransformation in Humans 7
2.2.3 Reactivity towards Biomacromolecules ...................................................... 8
2.2.4 Enzymes involved in Acrylamide Metabolism: .............................................
Function, Mechanism and Polymorphism ................................................ 10
2.3 Acrylamide Toxicity ....................................................................................... 21
2.3.1 Acute Toxicity ........................................................................................... 21
2.3.2 Neurotoxicity ............................................................................................. 21
2.3.3 Reproductive Toxicity ............................................................................... 22
2.3.4 Carcinogenicity .......................................................................................... 23
2.4 Human Exposure to Acrylamide ................................................................... 28
2.4.1 Biomarkers of Exposure ............................................................................ 28
2.4.2 Workplace Exposure .................................................................................. 31
2.4.3 Exposure to Acrylamide Residues from Polyacrylamides ........................ 31
2.4.4 Smoking ..................................................................................................... 32
2.4.5 Dietary Exposure ....................................................................................... 33
2.5 Formation of Acrylamide in Food ................................................................. 35
2.6 Risk Assessment .............................................................................................. 39
2.6.1 Human Risk Assessment of Genotoxic Carcinogens ................................ 39
2.6.2 Human Risk Estimations for Acrylamide Toxicity ................................... 42
2.6.3 Gaps and Limitations in Human Risk Assessment of Acrylamide ............ 45
II Table of Contents
3 Objectives ........................................................................................................................ 47
4 Analytical Method Development .................................................................................... 49
4.1 Introduction ..................................................................................................... 49
4.1.1 Analysis of Acrylamide-derived Metabolites in Urine ............................... 49
4.1.2 Hydrophilic Liquid Interaction Chromatography ....................................... 50
4.1.3 Electrospray Ionization ............................................................................... 54
4.1.4 Multiple Reaction Monitoring Mode .......................................................... 55
4.2 Materials and Methods ................................................................................... 56
4.2.1 Instrumentation ........................................................................................... 56
4.2.2 Chemicals ................................................................................................... 57
4.2.3 Synthesis of Glyceramide ........................................................................... 57
13 24.2.4 Synthesis of AAMA, C -AAMA and H -AAMA .................................. 58 3 3
134.2.5 Synthesis of AAMA-sulfoxide, C -AAMA-sulfoxide ................................ 3
2 and H -AAMA-sulfoxide .......................................................................... 58 3
134.2.6 Synthesis of C -Glycidamide ................................................................... 59 3
13 24.2.7 Synthesis of GAMA, C -GAMA and H -GAMA .................................. 60 3 3
4.2.8 Purification and Characterization of Synthesis Products ........................... 61
4.2.9 Liquid Chromatography ............................................................................. 63
4.2.10 Mass Spectrometry ..................................................................................... 65
4.2.11 Calibration Curves and Quantification ....................................................... 65
4.2.12 Method Validation ...................................................................................... 67
4.2.13 Human Study: Smokers vs. Nonsmokers ................................................... 67
4.2.14 Sample Preparation 68
4.2.15 Analysis of Creatinine ................................................................................ 68
4.3 Results and Discussion .................................................................................... 68
4.3.1 Synthesis of Glyceramide ........................................................................... 68
13 24.3.2 Synthesis of AAMA, C -AAMA and H -AAMA .................................. 69 3 3
134.3.3 Synthesis of AAMA-sulfoxide, C -AAMA-sulfoxide ................................ 3
2 and H -AAMA-sulfoxide .......................................................................... 70 3
134.3.4 Synthesis of C -Glycidamide ................................................................... 71 3
Table of Contents III
13 24.3.5 Synthesis of GAMA, C -GAMA and H -GAMA ................................. 72 3 3
4.3.6 Column Switching HILIC-MS/MS Method .............................................. 73
4.3.7 Method Validation ..................................................................................... 78
4.3.8 Analysis of AAMA, AAMA-sulfoxide and GAMA in Urine ................... 82
4.4 Conclusions ...................................................................................................... 83
5 Human Biomonitoring of Acrylamide Exposure ......................................................... 85
5.1 Introduction .................................................................................................... 85
5.2 Materials and Methods .................................................................................. 86
5.2.1 Human Study: Biomonitoring of Dietary Acrylamide Exposure .............. 86
5.2.2 Sample Preparation .................................................................................... 87
5.2.3 HILIC-MS/MS Analysis of AAMA, AAMA-sulfoxide and GAMA ........ 87
5.3 Results and Discussion 87
5.4 Conclusions ...................................................................................................... 93
6 Biotransformation and Toxicokinetics of Acrylamide in Rats ..................................... 95
6.1 Introduction .................................................................................................... 95
6.2 Materials and Methods .................................................................................. 96
6.2.1 Rat Study: Animals and Handling ............................................................. 96
6.2.2 Sample Preparation .................................................................................... 96
13 136.2.3 HILIC-MS/MS Analysis of C -AAMA and C -GAMA .......................... 3 3
in Rat Urine ............................................................................................... 97
6.2.4 Toxicokinetic Analysis .............................................................................. 98
6.3 Results and Discussion ................................................................................... 99
13 136.3.1 Urinary Excretion